This chapter focuses on the estimation of parameter affinity in rhizobacterial cocktail formulations for bio-recovery of crude oil-impacted soil. The topic relied on a study investigating the utilization of locally available substrates in ecologically disturbed ecosystems, with a focus on the rhizosphere of weeds growing on aged crude oil-impacted soil in the Niger Delta region. The identified rhizobacterial isolates: Achromobacter agilis, Pseudomonas fluorescens, Bacillus thuringiensis, and Staphylococcus lentus, are renowned for significant biodegradative potentials. The researchers assessed the impact of different parameters on growth dynamics of these isolates. By utilizing agro-residues like corn chaff as carbon source, corn steep liquor for nitrogen, and poultry droppings for phosphorus, as sources of limiting nutrients, the researchers varied factors like nutrient availability, pH levels, and temperature to estimate the affinity of these parameters for growth formulations and bioremediation capabilities by fitting the substrate utilization data into a Growth Kinetics Models. Data obtained revealed the isolates’ affinity for different substrates and provide valuable insights for optimizing the composition and performance of rhizobacterial cocktails for efficient hydrocarbon degradation in crude oil-impacted soil. Additionally, they underscored the potential of locally available substrates and microbial flora as effective tools for bio-recovery of crude oil-impacted soil.