In the last decades legal and other cases in which forensic veterinary science is required are increasing. The principles of investigation follow those used in human beings, although in animal species familial relationship identification is a hard task, due to high level of inbreeding. Familias 3 software is commonly used in human forensic laboratories to calculate probabilities and likelihood ratio (LR) in forensic cases. In the present paper we applied the same principles used in humans to standardize, simplify and accelerate the analysis, applying Familias 3 software in animals. Currently short tandem repeats identification is used as an effective method to trace DNA markers in genotyping. The authors used a standardized microsatellite panel recommended by International Society for Animal Genetics (ISAG) and a large allelic frequency Sicilian bovine DNA profiles database previously developed and published (Cosenza 2015). Based on this bovine matrix, the software permits the allelic frequency data calculation for the investigated samples and the data comparison to find the direct parentages and sibling. We analysed the most common veterinary forensic cases by using Familias 3 modules, alone or combined, to identify the proper module for each case. The software analysis was tested on 196 samples. A practical application of such system should be complementary to the conventional traceability, based on traditional labelling (Guhrin, 1994; Egeland, 2000). The impact of this software on the quality of the analysis report is critical to solve illegal inquiry, as much as the wrong calculation and data interpretation can invalidate the DNA profiling.