Bruises in pigs inflicted by blunt trauma are a significant animal welfare problem, and affected skin and underlying muscle are regularly submitted for forensic investigation. Central to the evaluation is an assessment of the age of the bruises. This paper presents cases of bruises in pigs sent for forensic investigation that were collected retrospectively. Data comprised photographs of the gross lesions, slides for histology, and written reports. The time from collecting the animals at the farms and delivery to the slaughterhouse was recorded together with the time of slaughter. Since 2005 there has been an increase in cases, with a peak in 2008 and 2009 of 40 cases for each year. At gross examination, the pattern of bruises often reflected the type of object which caused them. Histologically, haemorrhage and cellular infiltrations were frequently present. Currently, the age of bruises may be estimated to be more or less than four hours based on a porcine bruise model. In bruises more than four hours old, estimations of two-hour intervals are used based on studies of wound healing. The time from collecting the pigs at the farms until slaughter was between one and four hours in 44.1 per cent of cases, during which time the pigs had been handled by several people. In addition, in 22.0 per cent of cases of bruising an inflammatory response was absent, making it impossible to estimate the age of the bruise.

The model was reproducible and resembled forensic cases of bruises in pigs and humans. Therefore, the histological parameters are suitable for age determination of bruises in pigs and likely also in humans.

The assessment of the age of bruises inflicted on livestock is an important component of veterinary forensic pathology investigations. However, the sampling site within a bruise, the anatomical location and the mass and speed of the object inflicting the blunt trauma might influence the intensity of the inflammatory reaction. In the present study, the variation of the inflammatory reaction within and along experimental porcine bruises was evaluated in order to determine the optimal sampling site. Moreover, we evaluated if a combination of histological characteristics and gene expression signatures was able to differentiate bruises according to anatomical location, age of bruises and the speed and mass of the object used to cause the impact. Twelve experimental slaughter pigs were anesthetized, and on each animal four blunt traumas were inflicted on the back using either a plastic tube or an iron bar, respectively. The pigs were euthanized at 2, 5 or 8 h after infliction. Following gross examination, skin and underlying muscle tissue were sampled from the center and both ends of bruises and evaluated histologically. Subcutaneous fat tissue from the center of the bruises was sampled for quantitative real-time polymerase chain reaction to evaluate mRNA expression of 13 selected genes. Uninjured tissue was sampled from the right thigh of all pigs and served as control tissue. The amount of tissue damage and the intensity of the inflammatory reaction in bruises depended on the sampling site within and along a bruise, the anatomical location and the age of the bruise. The optimal site for sampling, i.e. the most pronounced inflammatory reaction, was at the center of the bruises where the plastic tube or iron bar first struck the skin. Moreover, bruises inflicted in areas with a thin layer of subcutaneous fat tissue showed more damage and inflammation in the underlying muscle tissue compared to bruises inflicted in areas with a thicker layer of subcutaneous fat tissue. In addition, hemorrhage in the muscle tissue was more likely present when bruises were inflicted with an iron bar compared to a plastic tube. Combining histology and mRNA expression of the 13 genes showed that the age of bruises could be determined with a precision of ±2.04 h. Moreover, the age of bruises could be determined with a precision of ±1.84 h based solely on mRNA expression of a selection of four genes.

Determining the age of bruises and the force used to inflict the trauma is of crucial importance in both human and veterinary forensic pathology. In the present study, the expression of more than 50 different genes in subcutaneous fat and muscle tissue from experimental bruises in pigs was investigated. The aim was to evaluate if expression signatures of selected genes were capable of determining bruises according to age and the force of impact. Eighteen experimental pigs were anesthetized, and on each animal four blunt traumas were inflicted on the back with a low, moderate or high force. The pigs were euthanized from 1 to 10 h after infliction of the trauma and subcutaneous fat and muscle tissues were sampled. As control, subcutaneous fat and muscle tissues were sampled from two un-injured pigs. Quantitative real-time polymerase chain reaction was performed to evaluate mRNA expression of genes involved in inflammation, tissue damage and repair. Expression signatures of thirteen selected genes in subcutaneous fat but not in muscle tissue reflected the age of bruises with a precision of approximately ±2 h. Moreover, the gene expression signature in the subcutaneous fat was to some extend able to separate bruises inflicted with different forces. Expression signatures of selected genes in the subcutaneous fat will increase the precision of the age determination of bruises in pigs. Further, due to the similarity of porcine and human skin physiology and immunity, these results might also provide valuable information in human forensic science.

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