Identification of Antemortem and Postmortem Injuries in Nude Mice Based on Microbial Communities

doi:10.12116/j.issn.1004-5619.2023.430806

Abstract

Abstract:

Objective To establish antemortem and postmortem injury models in nude mice, observe the morphological changes of the wounds and the changes of the microbial communities in the wounds at different time points after the injury, and analyze the differences between antemortem and postmortem wounds. Methods Abdominal injury models were established in 48 BALB/c nude mice, which were classified into antemortem injury, 4 h and 72 h postmortem injury groups, and the gross manifestations and histopathological changes were observed on days 1, 3, 5, 8, 11 and 15 after injury. The microbial communities in the wounds were analyzed by 16S rRNA sequencing technology. QIIME 2 software was used to calculate Shannon and Observed species indices. The Kruskal-Wallis test was used to determine statistical differences in α-diversity between groups. Jaccard similarity coefficients were calculated by using R v4.3.0 software and applied to the principal co-ordinates analysis to demonstrate inter-sample differences. Permutational multivariate analysis of variance （PERMANOVA） was used to analyze the differences between groups in the composition of bacterial colonies, and R² values were calculated. Results On days 8, 11 and 15 after injury, the antemortem and postmortem injuries could not be differentiated by morphological examination; the Shannon index and Observed species index were statistically different between the antemortem injury group and the 72 h postmortem injury group; the Jaccard similarity coefficient of the microbial community was statistically different between the antemortem injury group and the 72 h postmortem injury group. The PERMANOVA R² value gradually increased with the extension of time （0.22-0.61）. Conclusion Through the analysis of the wound microbial community, the microbial composition of wounds at different time points can be identified and compared, which provides a new perspective and method for the differentiation of antemortem injuries from postmortem injuries, with good application prospects.

Key words: forensic pathology, microbiology, colony, antemortem injury, postmortem injury, 16S rRNA, BALB/c nude mice

CLC Number:

Xin ZHENG, Yue QIU, Zhi-gang LI, Qing-qing XIANG, Guan-san WANG, He SHI, Qu-yi XU, Peng SUI, Yan-bing MA, Chao LIU, Li-fang CHEN, Jian ZHAO. Identification of Antemortem and Postmortem Injuries in Nude Mice Based on Microbial Communities[J]. Journal of Forensic Medicine, 2024, 40(5): 430-438.

Figures/Tables 7

Fig. 1 Three wound locations in the nude mouse

Fig. 2 Morphological changes of the wounds in 3 groups at different time points after injury

Fig. 3 Histopathological changes of the wounds in 3 groups at different time points after injury （HE×400）

Fig. 4 The distribution of the top 10 species in terms of relative abundanceat different time points after injury in 3 wound groups

Fig. 5 Results of alpha diversity analysis of 3 groups of traumatic bacterial communities

Fig. 6 PCoA results of 3 wound groups on days 8， 11， 15 after injury

Fig. 7 Screening results of differential species

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