Nanocrystalline NiFe2O4 particles prepared by chemical co-precipitation method were studied using magnetic measurements, 57Fe Mössbauer spectroscopy, X-ray diffraction, and transmission electron microscopy. Fits to Mössbauer spectra, in the range of 4.2 K – 300 K, were done using spin hamiltonians to describe both the electronic and nuclear interactions, a model of superparamagnetic relaxation of two levels (spin ½) and stochastic theory, a log-normal particle size distribution function as well as a dependency of the magnetic transition temperature and the anisotropy constant on particle diameter. We have used evolutionary strategies to fit the more complex Mössbauer spectra line shapes. The nanoparticles have an average size of 7 nm and exhibit superparamagnetism at room temperature. The saturation magnetization (Ms) at 4.2 K was determined from M vs. 1/H plots by extrapolating the value of magnetizations to infinite fields, to 24.21 emu/g and coercivity to 3.15 kOe. A magnetic anisotropy energy constant (K) 1.9´105 J/m3, at 4.2 K, were calculated from magnetization measurements. The synthesis, characterization, and functionalization of magnetic nanoparticles is a highly active area of current research located at the interface between materials science, biotechnology, and medicine. Superparamagnetic iron oxides nanoparticles have unique physical properties and have emerged as a new class of diagnostic probes for multimodal tracking and as contrast agents for magnetic resonance imaging (MRI).