Genetic analysis in animals has been used for many applications, such as kinship analysis, for determining the sire of an offspring when a female has been exposed to multiple males, determining parentage when an animal switches offspring with another dam, extended lineage reconstruction, estimating inbreeding, identification in breed registries, and speciation. It now also is being used increasingly to characterize animal materials in forensic cases. As such, it is important to operate under a set of minimum guidelines that assures that all service providers have a template to follow for quality practices. None have been delineated for animal genetic identity testing. Based on the model for human DNA forensic analyses, a basic discussion of the issues and guidelines is provided for animal testing to include analytical practices, data evaluation, nomenclature, allele designation, statistics, validation, proficiency testing, lineage markers, casework files, and reporting. These should provide a basis for professional societies and/or working groups to establish more formalized recommendations.

The present study investigated the capabilities and performances of semi-continuous and fully-continuous probabilistic approaches to DNA mixtures interpretation, particularly when dealing with Low-Template DNA mixtures. Five statistical interpretation software, such as Lab Retriever and LRmix Studio - involving semi-continuous algorithms - and DNA•VIEW, EuroForMix and STRmix- employing fully-continuous formulae - were employed to calculate likelihood ratio, comparing the prosecution and the defense hypotheses relative to a series of on-purpose prepared DNA mixtures that respectively contained 2 and 3 known contributors. National Institute of Standards and Technologies (NIST) certified templates were used for samples set up, which contained different DNA amounts for each contributor. 2-person mixtures have been prepared with proportions equal to 1:1, 19:1 and 1:19 in terms of DNA concentration. Conversely, three person mixtures were constituted by proportions equal to 20:9:1, 8:1:1, 6:3:1 and 1:1:1 in terms of DNA concentration. Furthermore, 8 equally-proportioned 3-person mixtures were prepared by means of scalar dilutions starting from an overall amount of 0.500 ng, then ranging up to DNA samples with concentrations equal to 0.004 ng (i.e. Low-Template DNA). DNA mixtures were set up in triplicate and amplified with 7 DNA amplification kits (i.e. GlobalFiler PCR Amplification Kit, NGM SElect PCR Amplification Kit, MiniFiler PCR Amplification Kit, Power Plex Fusion, PowerPlex 6C Matrix System, Power Plex ESI 17 Fast and Power Plex ESX 17 Fast) in order to evaluate whether the selection of a certain kit might represent a bias factor, capable of altering the whole interpretation process. A multi-software approach helped us to highlight any trend in the likelihood ratio results provided by semi- and fully-continuous software. As a matter of fact, fully-continuous computations provided different (higher) results in terms of degrees of magnitude of the likelihood ratio values with respect to those from the semi-continuous approach, regardless of the amplification kit that was utilized.

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