Analysis of microsatellite polymorphism in red deer, roe deer, and fallow deer -- possible employment in forensic applications

DNA microsatellites play a major role in population genetics, linkage mapping, and parentage studies of mammals. In addition, they may be used for forensic purposes, if an individual identification of a specific animal is necessary. Therefore, we tested a variety of microsatellite polymorphism derived from reindeer (Rangifer tarandus) by PCR and sequencing analysis for use in red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama). Twelve of these microsatellites were selected for further analysis. In all these microsatellite polymorphism short tandem repeats could be detected for one or all three species as shown by sequencing analysis. In red deer, more than two alleles were found in eight microsatellites, in roe deer more than two alleles could be demonstrated in seven microsatellites, whereas in fallow deer more than two alleles were found in only two microsatellite polymorphism. A comparison of sequences of PCR products from the three deer species with the sequences of reindeer revealed several differences between the four species. In six microsatellites -- selected because or their reliability in PCR and because of their polymorphic character -- we established a sequenced allelic ladder and give population data of all three species from 82 deer of the Northeast region of Germany (Vorpommern). Our results show the possibility to use microsatellite polymorphism in the identification of deer in forensic applications like poaching.     

Sex identification of white-tailed deer using frozen venison

Fluids from frozen muscle tissue taken from 449 white-tailed deer (Odocoileus virginianus) were analyzed using a radioimmunoassay technique for testosterone. Comparing 309 deer sampled in early November, antlered males had significantly higher tissue testosterone levels (range = 0.18 to 16.00 ng/mL) than females (range = 0.00 to 0.18 ng/mL) while antlerless (fawn) males overlapped both groups (range = 0.00 to 0.68 ng/mL). Samples taken from 132 deer from January through August showed no differences among ages or sexes. Antlered males can be differentiated from females during the breeding season.     

DNA forensics and the poaching of wildlife in Italy: a case study

DNA molecular techniques were used in a forensic investigation involving the poaching of wildlife in a national park of Italy. A poacher, after having snared a wild boar (Sus scrofa) sow, knifed it to death. The animal was retrieved by conservation officers at the scene before the poacher could remove the carcass. Subsequently, the suspect denied the charges. During a search of his home, a bloodstained knife was confiscated. A method to identify the species from the DNA extracted from the stains revealed the blood to be that of the non-domestic form of Sus scrofa. Further DNA typing for individual identity using species-specific single tandem repeats or microsatellites (STRs) showed that the DNA on the knife matched that of the poached boar. Based upon the forensic evidence obtained, the suspect was convicted of poaching and of cruelty to animals.     

A molecular genetic approach for species identification of mammals and sex determination of birds in a forensic case of poaching from South Korea

DNA-based analysis was performed using partial mitochondrial cytochrome b genes of five mammalian specimens and Chromo-Helicase-DNA-binding (CHD) genes of five pheasants to determine whether specimens were from illegally hunted animals. Mammalian specimens were identified as being those of horse, roe deer, and cow through gene amplification using cytb981f and cytb981r primer set and sequencing. CHD genes were revealed to be those of three male and two female pheasants through polymerase chain reaction amplification. Because hunting of roe deers and female pheasants is prohibited in Korea, these results provided forensic evidences of illegal wild animal hunting.     

Individual identification of disguised faces by morphometrical matching

The reliability of a morphometrical matching method for identifying disguised faces was examined experimentally using a computer-assisted facial image identification system. The 2D right oblique facial images of three target persons disguised with sunglasses, cap and gauze mask were each compared with each of the 3D facial images of 100 subjects, yielding 900 face-to-face superimpositions. The average perpendicular distance between the facial outlines and the average point-to-point distance of the corresponding landmarks in the 2D image of the disguised face and the 3D facial image, were calculated. As a matching criterion, the sum of the values of the average perpendicular difference of the facial outlines and the average point-to-point difference between the corresponding landmarks was used (abbreviation: average difference).The range of the average difference was 2.3-2.8mm for the same person (a match) and 4.0-14.6mm for different (non-matching) people, respectively. The ranges for matching and non-matching faces did not overlap. Even the 3D facial images of the non-matching person showing the closest value of average difference to the average difference for the matching person could be excluded easily. It was concluded that the morphometrical matching method can reliably identify disguised faces and the results produced by this method could be easily understandable by a court of law.     

Forensic DNA against wildlife poaching: Identification of a serial wolf killing in Italy

The recent expansion of the Italian wolf population through the Apennine and western Alps, after centuries of contractions, is causing conflicts with human activities leading to a rise in poaching or illegal killings. Here we show how molecular population genetics has been used to identify a suspect serial wolf killer. We analysed DNA extracted from a necklace made of ten presumed wolf canine teeth, confiscated in 2008 to a man living in the northern Italian Apennine (Liguria Region). Individual genotypes were determined using 12 unlinked autosomal microsatellites (STRs), mtDNA control-region sequences, a male-specific ZFX/ZFY restriction-site and three Y-linked STRs. Results indicate that the teeth belonged to six different individuals (three males and three females), which were assigned to the Italian wolf population with p > 0.90 by Bayesian procedures. One of these genotypes matched with the genetic profile of a male wolf previously found-dead and already non-invasively sampled in the same area. Another genotype matched with that of a female wolf non-invasively sampled twice in the same area 1 year before. These data are being used as forensic genetic evidence in the ongoing criminal trial against the suspect serial wolf killer.     

The polymorphism of alpha-amylase]

Individual phenotypes, phenotypic and genetic frequencies of alpha-amylase enzyme were determined by means of population genetic study. The results of this study revealed absence of genetic linkage between alpha-amylase phenotypes, haptoglobins and serum factor G1m(1) of gammaglobulin system (Gm).     

mtDNA多态性在嗜尸性昆虫种类鉴定中的应用

嗜尸性昆虫的种类鉴定是利用昆虫学知识进行法医学研究的重要步骤之一.近年来研究表明,利用分子生物学技术,特别是线粒体DNA(mitochondrial DNA,mtDNA)序列分析技术对嗜尸性昆虫进行种类鉴别,能够得到较好的效果.本文对嗜尸性昆虫mtDNA的分子生物学特性、结构以及嗜尸性昆虫种类鉴定中常用的mtDNA序列...     

唾液酯酶多态性在个体识别中的应用研究

目的 探讨唾液酯酶（ Set）多态性在法医学亲权鉴定和个体识别方面的应用价值。方法 应用聚丙烯酰胺凝胶盘状电泳及固蓝 RR染色方法,调查了 114名中国人 Set的表型分布及基因频率,用χ 2检验进行统计学分析。结果 中国人酯酶表型频率 Set F 22.81％, Set FS 50.88％, Set S 26.31％ ;基因频率为 SetF 0.482 5, SetS 0.517 5;非父排除机率为 0.187 5,个体识别率为 0.619 9。结论 Set有较高的父权排除率和个体识别率,可作为法医学亲权鉴定和个体识别的重要标记系统之一。     

Application of mitochondrial DNA sequence analysis in the forensic identification of Chinese sika deer subspecies

As a direct and indirect consequence of human activities, only two subspecies, Cervus nippon sinchuanicus and Cervus nippon kopschi, currently subsist in the wild of China. However, a large population of Cervus nippon hortulorum and Cervus nippon nippon is raised in order to gain deer parts for Chinese traditional medicine. According to Chinese Wild Animal Conservation Law, hunting, capturing and trading of the wild sika deer are strictly banned, however, raising and trading of the domestic individual are permitted. Thus, it is very necessary to identify the subspecies of sika deer in China in forensic tests. In our study, we used mitochondrial DNA control region sequence analysis and phylogenetic analysis to identify the subspecies of sika deer. Mitochondrial DNA control region sequences analysis revealed that two haplotypes came from the unknown samples. One is the same as the haplotype that came from the samples of wild population of C. n. kopschi. Phylogenetic analysis indicated that the two haplotypes of unknown samples clustered with the haplotypes of C. n. kopschi, and had significant difference from the haplotypes of the other subspecies. These results together revealed that the unknown samples came from two individuals that belong to the wild population of C. n. kopschi living in the Qinglingfeng State Natural Reserve of Zhejiang province. Therefore, the results provide forensic evidence of illegal wild animal hunting.     

Genetic polymorphism at nine microsatellite loci in four high altitude Himalayan desert human populations

Allele frequencies for the nine STR loci of Profiler Plus STR markers were analyzed in 307 healthy unrelated individuals belonging to four predominant highly important endogamous population groups of Ladakh. The studied loci are found highly informative in human identification and understanding history of peopling of India. This is the first report on DNA Markers on any high altitude desert human population group.     

应用AFLP检测大麻遗传多样性

目的 筛选对于大麻多态性好的AFLP引物标记来区分大麻品种.方法 利用AFLP技术用55对引物组合对12个大麻地域品种进行初筛.结果 选出5对多态性好的引物组合进行了遗传多样性研究.每对AFLP引物组合扩增出47～76务带.共获得285条带,其中多态性条带为99条以及10条品种特异带.结论 AFLP对于大麻具有很高的分辨率,为今后深入地研究大麻植物遗传多样性奠定了很好的基础.     

Forensic identification of a rapist using unusual evidence.

This case report demonstrates a rape case, where no semen, hair, or fingerprints were left by the perpetrator at the crime scene, but rather uncharacteristic biological and physical evidence in the form of a lollipop and a pair of glasses. Three separate forensic laboratories collaborated using conventional forensic methods of PCR DNA typing, photography, and toolmark comparisons to provide investigators with scientific evidence which in turn was instrumental in bringing a violent criminal to justice. The importance of evaluating each item of evidence and realizing its forensic value is stressed in this case report.     

Bullet identification: a case of a fatal hunting accident resolved by comparison of lead shot using instrumental neutron activation analysis.

Bullet identification by chemical analysis often provides a powerful clue in forensic science. A case is reported in which a hunting accident was resolved by using instrumental neutron activation analysis (INAA) for direct comparison of the trace element content in lead shot. Different preparation batches of lead shot appear to have a high within-group composition homogeneity, and good differentiation is achieved between different batches. Determination of the nickel and antimony content on a bush branch demonstrated that the branch had been perforated by one of the shot pellets, and this helped the detectives in reconstruction of the crime scene.     

Human genotyposcopy: the identification of the species, sex and individual by DNA genetic imprints in a case connected with a murder attempt]

Blood stains on a knife were identified by DNA genotyposcopy. The statistical validation method has confirmed that the blood stains on material evidence belonged to the victim, the probability of random coincidence being less than 10(-11). The efficacy of using hypervariable locus-specific DNA probes and the possibility of detecting DNA impressions in blood stains stored for more than 3 months have been demonstrated.     

SNPs在个体识别与表型预测中的研究进展

单核苷酸多态性（single nucleotide polymorphism,SNP）作为第三代遗传标记,具有分布广泛、突变率低、遗传稳定及易于自动化高通量快速检测分析的特点。同时,因其扩增片段长度短、不存在复制滑动,所以利于腐败降解、痕量检材的检测。随着研究的深入,SNPs在法医学领域受到了广泛重视,与表型（ABO血型、色素沉积及颅面形态）相关的SNPs有望用于预测嫌疑人的基本特征,为案件侦破提供新的思路。本文对近年来SNPs在个体识别和表型预测的研究进行总结,介绍该领域的研究进展,为法医学工作者提供参考。     

Weapon identification using antemortem computed tomography with virtual 3D and rapid prototype modeling-A report in a case of blunt force head injury

This research shows the exfoliation of the anterior dentition has significant potential to aid in establishing the minimum length of the post-mortem interval. Accumulated degree days (ADD) were used to quantify the decomposition of the periodontal ligament, represented by post-mortem exfoliation of the incisors. After subjects were removed subsequent to disturbance by scavengers and time limitations on the study, the final sample size was 36 incisors from the maxillae and mandibles of seven pigs (Sus scrofa). Average daily temperature was calculated using hourly temperature data recorded using DS1921G thermochrons for the duration of the project (June 14-December 17, 2008). During this period, six teeth (16.7%) were exfoliated. ADD for these six teeth ranged from 1539.7 °C to 2006.7 °C. The average ADD required for exfoliation was 1788.0 °C (SD=198.1 °C). No differences in ADD required for exfoliation were observed between the maxillary and mandibular teeth (t=2.085; p=0.128).