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BM2: Council Information BM2:理事会信息
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2026-01-01 DOI: 10.1016/S1355-0306(25)00173-X
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引用次数: 0
Encapsulation of degraded DNA in alginate hydrogels: Rheological characterization and applicability to forensic science 海藻酸盐水凝胶中降解DNA的包封:流变学表征及其在法医学上的适用性
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-26 DOI: 10.1016/j.scijus.2025.101378
Colin I. Elliott , Theresa Stotesbury
Forensic biomaterials are on the rise, with efforts focused on developing realistic tissue and blood mimetics. The incorporation of small and degraded DNA into these materials enhances their realism and functionality, which has implications for research and training across forensic science. It is therefore important to understand the physicochemical and conformational changes that DNA undergoes during ex vivo degradation. Large fragments of highly concentrated genomic and phage DNA in solution have been characterized using rheology; however, this amount and size of DNA are atypical in DNA extracted from forensic evidence. In this work, we investigated how the addition of synthetic DNA oligos and genomic DNA extracted from bloodstains deposited for up to 19 months influenced the rheological properties of polymer systems intended for forensic biomaterial synthesis. We used FTIR spectroscopy to probe interactions between DNA and the encapsulating matrix and automated gel electrophoresis to record DNA quality/quantity metrics, both of which supported our rheological findings. Encapsulating DNA within an alginate-based, ionically crosslinked hydrogel produced the greatest differentiation in rheological profiles among DNA with varying physical properties. The distinct conformations and sizes of encapsulated DNA oligos exhibited significantly different responses during strain amplitude sweeps (p < 0.05). We also observed moderate correlations between the rheological responses of DNA extracts and the time since deposition of corresponding bloodstains (r = −0.57 to r = 0.62). This indicates that dilute, polydisperse and degraded genomic DNA extracts can modulate the rheological properties of the encapsulating hydrogel, highlighting the need to consider the type of DNA included in forensic biomaterials. Our results demonstrate the potential for rheology to serve as a complementary technique when analyzing encapsulated dilute DNA oligos and degraded DNA.
法医生物材料正在兴起,重点是开发逼真的组织和血液模拟物。将小的和降解的DNA结合到这些材料中,增强了它们的真实感和功能,这对法医科学的研究和培训具有重要意义。因此,了解DNA在体外降解过程中所经历的物理化学和构象变化是很重要的。用流变学方法对溶液中高浓度基因组和噬菌体DNA的大片段进行了表征;然而,这种数量和大小的DNA在从法医证据中提取的DNA中是不典型的。在这项工作中,我们研究了从沉积长达19个月的血迹中提取的合成DNA寡核苷酸和基因组DNA的添加如何影响用于法医生物材料合成的聚合物系统的流变性能。我们使用FTIR光谱来探测DNA与封装基质之间的相互作用,并使用自动凝胶电泳来记录DNA的质量/数量指标,这两者都支持我们的流变学研究结果。将DNA封装在海藻酸盐基的离子交联水凝胶中,在具有不同物理性质的DNA中产生了最大的流变谱差异。不同形状和大小的DNA寡核苷酸在应变振幅扫描过程中表现出显著不同的反应(p < 0.05)。我们还观察到DNA提取物的流变反应与相应血迹沉积时间之间存在适度的相关性(r = - 0.57至r = 0.62)。这表明,稀释、多分散和降解的基因组DNA提取物可以调节包封水凝胶的流变特性,强调需要考虑法医生物材料中包含的DNA类型。我们的研究结果表明,流变学在分析封装的稀释DNA寡聚物和降解DNA时,可以作为一种补充技术。
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引用次数: 0
Missing persons response: national programs and global cooperation in Brazil’s migration context 失踪人口应对:巴西移民背景下的国家方案和全球合作
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-21 DOI: 10.1016/j.scijus.2025.101379
Aline Minervino , António Amorim , Francisco Corte Real , Laura Cainé
The search and identification of missing persons represent a pressing global issue with profound humanitarian, legal, and institutional implications. This study presents an illustrative documentary review of MP/UHR identification initiatives in countries selected for their migration-related links to Brazil and their relevance to transnational DNA cooperation (Brazil, Portugal, Spain, France, the United Kingdom, the United States, Canada, Mexico, and Belgium). Grounded in the United Nations guidelines and using peer-reviewed literature, official reports, and documentation from international organizations, the analysis examine how forensic genetics is integrated into MP/UHR workflows, the legal and governance frameworks that enable or constrain the processing of relatives’ reference profiles, and the operational arrangements that support cross-border exchange. The synthesis indicates that identification outcomes depend not only on laboratory capacity, but also on institutional coordination and the linkage of ante-mortem and post-mortem data. The study also describes international mechanisms such as INTERPOL’s I-Familia database, and humanitarian identification platforms. Although these tools show great potential, their effectiveness is limited due to inconsistent adoption across countries and variations in national protocols. The identification of missing persons must be approached as a shared international responsibility, requiring cooperation in science, law, and policy to ensure dignity, truth, and justice for families of the missing.
失踪人员的搜寻和识别是一个紧迫的全球性问题,具有深刻的人道主义、法律和制度影响。本研究提供了一份说明性的文献综述,介绍了一些国家因其与巴西的移民相关联系以及与跨国DNA合作的相关性而选定的MP/UHR鉴定倡议(巴西、葡萄牙、西班牙、法国、英国、美国、加拿大、墨西哥和比利时)。该分析以联合国指导方针为基础,利用同行评审的文献、官方报告和国际组织的文件,研究了法医遗传学如何融入法医基因学/人权高专办工作流程,支持或限制亲属参考资料处理的法律和治理框架,以及支持跨境交流的业务安排。综合结果表明,鉴定结果不仅取决于实验室能力,还取决于机构协调以及死前和死后数据的联系。该研究还描述了国际机制,如国际刑警组织的I-Familia数据库和人道主义身份识别平台。尽管这些工具显示出巨大的潜力,但由于各国采用的不一致以及各国议定书的差异,它们的有效性受到限制。失踪人员的身份识别必须作为一项共同的国际责任来对待,需要在科学、法律和政策方面进行合作,以确保失踪人员家属的尊严、真相和正义。
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引用次数: 0
NIST recommended square footage for forensic laboratories compared with actual facilities: Size matters NIST建议将法医实验室的面积与实际设施进行比较:大小很重要
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-14 DOI: 10.1016/j.scijus.2025.101376
M.M. Houck , P.J. Speaker , A. Tessarolo
Trends in modern forensic laboratory construction reveal a significant increase in square footage per staff member compared to older facilities. This expansion stems from a blend of operational, regulatory, and architectural factors, rather than solely sustainability goals. The growing scientific scope, more rigorous accreditation standards, and the critical need for flexibility and secure evidence handling all contribute to these rising spatial requirements. While high-performance building frameworks like LEED can introduce additional design considerations, particularly for mechanical systems and circulation, sustainability is best understood as one influence among many. This technical note, drawing on Project FORESIGHT benchmarking data and NIST facility planning guidance, outlines the primary drivers of laboratory size in new construction and offers practical recommendations for facility planners aiming to align space allocation with forensic function.
现代法医实验室建设的趋势表明,与旧设施相比,每个工作人员的平方英尺显着增加。这种扩展源于运营、监管和建筑因素的混合,而不仅仅是可持续性目标。不断扩大的科学范围,更严格的认证标准,以及对灵活性和安全证据处理的迫切需求,都促成了这些不断增长的空间需求。虽然像LEED这样的高性能建筑框架可以引入额外的设计考虑,特别是在机械系统和循环方面,但可持续性最好被理解为众多影响中的一个。本技术说明借鉴了Project FORESIGHT基准数据和NIST设施规划指南,概述了新建实验室规模的主要驱动因素,并为旨在使空间分配与法医功能保持一致的设施规划者提供了实用建议。
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引用次数: 0
Shored bullet entry wounds: key practical insights in aid of shooting investigations 支撑子弹射入伤:帮助射击调查的关键实践见解
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-12 DOI: 10.1016/j.scijus.2025.101375
Bandula Nishshanka , Chris Shepherd , Randika Ariyarathna
“Foreshoring” is a rarely examined phenomenon in shooting incidents and occurs when a bullet traverses an intermediate surface that is firmly pressed against the body prior to skin penetration. Building upon the only experimental study available to date, published in 1980, this empirical investigation systematically evaluated the macroscopic features of shored bullet entry wounds produced using a range of commonly encountered shoring materials and bullet types relevant to forensic casework. Porcine skin samples mounted in a custom-designed experimental apparatus were used to generate shored entry wounds, which were subsequently compared with control wounds created in the absence of a shoring surface. The influence of varying downward forces on wound morphology was also assessed. Statistically significant macroscopic differences between shored and standard bullet entry wounds have been identified, revealing novel wound profiles and underlying mechanisms. Findings demonstrate that downward force does not alter the macroscopic morphology of shored-entry wounds, and even minimal separation between the skin and the supporting surface produces wound characteristics distinct from those of true shoring. The study also clarifies previous misconceptions, including a masking effect in close-range shots where gunshot residue may obscure key wound features. While presenting novel insights concerning shored gunshot entry wounds for practical forensic caseworks, the findings of this study highlight that the accurate recognition of shored-entry wounds is essential for reconstructing shooting incidents, interpreting wound patterns, and preventing misclassification in medicolegal investigations.
在射击事件中,“前冲”是一种很少被研究的现象,发生在子弹在穿透皮肤之前穿过紧贴身体的中间表面时。在1980年发表的迄今为止唯一可用的实验研究的基础上,本实证研究系统地评估了使用一系列常见的支撑材料和与法医案件相关的子弹类型产生的支撑子弹射入伤的宏观特征。将猪皮肤样本安装在定制设计的实验装置中,用于产生有支撑的进入伤口,随后将其与没有支撑表面的对照伤口进行比较。不同的向下作用力对创面形态的影响也进行了评估。统计上显著的宏观差异在支撑和标准子弹射入伤口之间已经被确定,揭示了新的伤口特征和潜在的机制。研究结果表明,向下的力不会改变支撑伤口的宏观形态,即使皮肤和支撑表面之间的最小分离也会产生与真正支撑伤口不同的伤口特征。该研究还澄清了之前的误解,包括近距离射击时的掩蔽效应,其中射击残留物可能会掩盖关键的伤口特征。本研究的结果在为实际法医案件提供有关枪弹射入伤的新见解的同时,强调了准确识别枪弹射入伤对于重建枪击事件、解释伤口模式和防止法医调查中的错误分类至关重要。
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引用次数: 0
Alginate-based forensic blood substitutes mimicking whole blood clotting and drying properties in drip stains and pools 海藻酸盐为基础的法医血液替代品模仿全血凝结和干燥特性滴渍和池
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-10 DOI: 10.1016/j.scijus.2025.101374
Erin Giroux , Shaijieni Kannan , Theresa Stotesbury , Mark C. Jermy
Forensic Blood Substitutes (FBS) may be used in place of blood for training and research in bloodstain pattern analysis. They offer a longer shelf life and less stringent biohazard precautions than animal or human blood. The drying dynamics of FBS have not been previously investigated. This preliminary work compares the drying dynamics of two alginate-based hydrogel FBS formulations with whole bovine blood deposited as drip stains and pools on glazed tile and cotton jersey fabric with pools of 2, 10 and 500 µL of each liquid. The first formulation (FBS1), previously optimized and validated for impact and drip patterns, was modified by adjusting the ionic crosslinker (CaCl2) and polymer (alginate) concentrations to create a second formula (FBS2) with more time-dependent viscosity to mimick the increasing viscosity of whole human blood during clotting. Rheometry was characterised with oscillation amplitude sweeps (0.1–150 % strain), flow sweeps (1–1200 s−1) and time sweeps (3.5 % strain and 1 Hz for 2000 s). In single tests, blood pools formed from FBS2 appear to show drying characteristics closer to whole blood than FBS1. On non-porous tile, the FBS materials broadly replicated the speed and appearance of the early stages of drying of blood. It developed radial stress lines in the corona, but did not crack in the way blood does in the final stages of drying. It did not de-bond from the tile surface in the same way. In the limited tests performed, FBS1 appears to spread further than blood to form larger stains, whereas FBS2 may not. On cotton jersey, both FBSs wicked at a similar rate to blood. Both FBS1 and FBS2 dried lighter than blood, but with the same dark edge to the stain. Ultimately, we found that by changing the polymer and ionic crosslinker concentration, the physical fluid properties of the material can be manipulated to behave more like blood during stain formation. Thus, the current formulations of the FBS are suitable for the generation of blood stains for training and research, with FBS1 acting as an accurate mimetic of whole blood for drip stains, and FBS2 possessing the viscous behavior required to better mimic whole blood clotting in larger volume stains and pools. Further work with more replicates, applied volumes and substrates is needed.
法医血液替代品(FBS)可以代替血液用于血迹模式分析的培训和研究。与动物或人类血液相比,它们具有更长的保质期和更宽松的生物危害预防措施。FBS的干燥动力学以前没有研究过。本初步工作比较了两种海藻酸盐基水凝胶FBS配方的干燥动力学,其中全牛血液作为滴痕和池沉积在釉面砖和棉织物上,每种液体的池分别为2,10和500 μ L。第一种配方(FBS1)之前针对冲击和滴注模式进行了优化和验证,通过调整离子交联剂(CaCl2)和聚合物(海藻酸盐)的浓度进行了修改,创造了第二种配方(FBS2),具有更多的时间依赖性粘度,以模仿凝血过程中全血粘度的增加。流变学的特点是振荡幅度扫描(0.1 - 150%应变),流量扫描(1 - 1200 s−1)和时间扫描(3.5%应变和1 Hz 2000 s)。在单次测试中,由FBS2形成的血池似乎比FBS1更接近全血的干燥特征。在无孔瓷砖上,FBS材料大致复制了血液干燥早期阶段的速度和外观。它在日冕上形成了径向应力线,但没有像血液在干燥的最后阶段那样破裂。它不会以同样的方式从瓷砖表面脱粘。在有限的测试中,FBS1似乎比血液扩散得更远,形成更大的污渍,而FBS2可能不会。在棉质运动衫上,这两种fbs的腐蚀速率与血液相似。FBS1和FBS2干后都比血淡,但有相同的深色边缘。最终,我们发现,通过改变聚合物和离子交联剂的浓度,可以操纵材料的物理流体特性,使其在染色过程中表现得更像血液。因此,目前的FBS配方适合用于训练和研究的血渍生成,FBS1可以准确模拟滴痕中的全血,而FBS2具有所需的粘性行为,可以更好地模拟大容量血渍和池中的全血凝固。需要对更多的复制、应用体积和底物进行进一步的研究。
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引用次数: 0
A bibliometric analysis of detection methods for clandestine graves in the context of human rights violations 在侵犯人权的情况下对秘密坟墓的侦查方法进行文献计量分析
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-06 DOI: 10.1016/j.scijus.2025.101373
Maira García-Jaramillo , Carlos Martín Molina , Fabian León-Vargas , Leonardo Torres , Alejandra Baena
The detection of clandestine graves is a critical task in forensic science, particularly in contexts of armed conflict, human rights violations, and organised crime. This study presents a bibliometric analysis of the scientific literature on clandestine grave detection published between 1996 and 2024, using Scopus as the primary data source. A total of 187 peer-reviewed publications were analysed through bibliometric indicators and visualization techniques, including co-occurrence and network analysis. The most prolific authors, institutions, countries, journals, and thematic trends were identified. Geophysical methods, particularly Ground Penetrating Radar and Electrical Resistivity Tomography, alongside forensic approaches such as volatile organic compounds detection and drone technology, have emerged as key techniques. Collaboration analysis highlights strong international networks, notably involving Keele University in the United Kingdom and several Latin American universities. However, regional disparities and underrepresentation of some conflict-affected regions persist in the literature. The study concludes by identifying research gaps and proposing directions for future interdisciplinary research.
发现秘密坟墓是法医学的一项关键任务,特别是在武装冲突、侵犯人权和有组织犯罪的背景下。本研究以Scopus为主要数据来源,对1996年至2024年间发表的关于秘密坟墓探测的科学文献进行了文献计量学分析。通过文献计量指标和可视化技术,包括共现分析和网络分析,共分析了187份同行评议出版物。确定了最多产的作者、机构、国家、期刊和主题趋势。地球物理方法,特别是探地雷达和电阻率层析成像,以及挥发性有机化合物检测和无人机技术等法医方法,已经成为关键技术。合作分析强调了强大的国际网络,特别是涉及英国基尔大学和几所拉丁美洲大学。然而,文献中仍然存在区域差异和一些受冲突影响地区的代表性不足。该研究通过确定研究差距并提出未来跨学科研究的方向来结束。
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引用次数: 0
Automated microscopy in forensic biology: Validating the use of deep neural networks for the detection of human spermatozoa in Christmas Tree stained slides 自动显微镜在法医生物学:验证使用深度神经网络检测人类精子在圣诞树染色玻片
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-04 DOI: 10.1016/j.scijus.2025.101370
Dean C. Topping, Teresa Tran, Colby M. Hymus, Nicholas S. Mountford, Yoon C. Liew, Scott E. Egan, Penny L. Cooper
The most widely accepted technique for the confirmation of semen presence in forensic biology is the microscopic identification of sperm cells. The process of manual microscopic examination, however, can often be time consuming, particularly where sperm numbers are low or where sperm cells are absent, particularly when in the presence of numerous non-sperm cells. The findings of this study demonstrate that automated microscopy utilising Deep Neural Network (DNN) allows for an improvement of the efficiency of examination times associated with microscopy. Additionally, the use of instrument and slide data as diagnostic tools permits rapid detection of invalid results. Metafer with Sperm Detection, using DNN v3.1.1, successfully identified sperm cells present in both high and low semen concentrations in mixed-cell substrates, often with a higher accuracy than manual microscopic examinations where very low sperm numbers were present (1–5 sperm cells on an entire slide). Whilst there were instances of false positive and false negative sperm cell classifications observed, the overall sensitivity, specificity and accuracy of automated microscopy using Metafer with Sperm Detection (DNN v3.1.1) was 0.9999, 0.9996 and 0.9999, respectively. The conversion of Metafer sperm counts to an existing sperm grading system allows compatibility with manual microscopy outcomes and allows decisions regarding sample triage and submission for specific DNA extraction protocols in routine casework to be made with confidence.
在法医生物学中,最广泛接受的确认精液存在的技术是精子细胞的显微鉴定。然而,人工显微镜检查的过程往往是耗时的,特别是在精子数量少或精子细胞缺失的情况下,特别是在存在大量非精子细胞的情况下。本研究的结果表明,利用深度神经网络(DNN)的自动显微镜可以提高与显微镜相关的检查时间的效率。此外,使用仪器和载玻片数据作为诊断工具,可以快速检测无效结果。Metafer with Sperm Detection使用DNN v3.1.1,成功地鉴定出混合细胞基质中精液浓度高和浓度低的精子细胞,通常比人工显微镜检查的准确性更高,其中精子数量非常低(整个载玻片上有1-5个精子细胞)。虽然观察到存在假阳性和假阴性精子细胞分类的情况,但使用Metafer with sperm Detection (DNN v3.1.1)的自动显微镜的总体灵敏度、特异性和准确性分别为0.9999、0.9996和0.9999。将Metafer精子计数转换为现有的精子分级系统,允许与人工显微镜结果兼容,并允许在常规病例工作中对样本分类和提交特定DNA提取方案做出有信心的决定。
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引用次数: 0
Effects of time, high ambient temperature, swab type, and preservatives on forensic DNA recovery 时间、高温、拭子类型和防腐剂对法医DNA恢复的影响
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-03 DOI: 10.1016/j.scijus.2025.101372
Maria Divya Sahayaselvan , Shruti Chowdhari , Vivekanandan Perumal , Shreya Rastogi
DNA obtained from biological samples plays a significant role in both criminal and civil investigations. However, the probative value of DNA evidence is heavily influenced by the conditions under which the biological samples are collected, handled, and preserved, given their susceptibility to degradation over time. While general guidelines for handling different types of biological samples have been established by the government agencies and forensic laboratories, specific procedures for handling biological samples intended for forensic DNA analysis remain poorly defined in developing countries. As unforeseen delays between sample collection and DNA testing are common in forensic settings, factors such as traditional collection methods, inadequate air drying prior to packaging, and storage in evidence rooms lacking temperature control often result in laboratories receiving poor-quality samples, which further hinders the efficiency of forensic DNA analysis. In this study, we demonstrate the influence of collection method, storage medium and duration of storage under high ambient temperature on DNA recovery. Our results show that (a) nylon-flocked swabs facilitate improved DNA yields, (b) the use of DNA-stabilizing preservatives (commercially available or prepared in-house) increases the amplifiable DNA yield of samples stored at 35 °C for up to one month, (c) no amplifiable DNA was recovered from either type of swabs when stored dry at high ambient temperature for more than a week without a preservative, and (d) regardless of the swab material and the use of preservative, no amplifiable DNA could be extracted from samples stored up to two months at 35 °C. In sum, this work demonstrates that the use of nylon-flocked swabs together with DNA-stabilizing preservatives has a tangible impact on the amount of amplifiable DNA recovered from samples stored at high ambient temperature for up to a month, and that incorporating these methods hold considerable forensic utility.
从生物样本中获得的DNA在刑事和民事调查中都起着重要作用。然而,DNA证据的证明价值在很大程度上受到生物样本的收集、处理和保存条件的影响,因为它们容易随着时间的推移而降解。虽然政府机构和法医实验室已经制定了处理不同类型生物样本的一般准则,但在发展中国家,处理用于法医DNA分析的生物样本的具体程序仍然没有明确规定。由于在法医环境中,样品收集和DNA检测之间的不可预见的延迟是常见的,传统的收集方法、包装前不充分的风干以及在缺乏温度控制的证据室中储存等因素往往导致实验室收到质量差的样品,这进一步阻碍了法医DNA分析的效率。在本研究中,我们展示了收集方法、储存介质和高温下储存时间对DNA恢复的影响。我们的研究结果表明:(a)尼龙棉签有助于提高DNA产量,(b)使用DNA稳定防腐剂(市售或自制)增加了在35°C下储存长达一个月的样品的可扩增DNA产量,(C)在没有防腐剂的情况下在高温下干燥储存超过一周时,两种棉签都没有从可扩增DNA中恢复,以及(d)无论棉签材料和防腐剂的使用如何。在35°C下储存两个月的样品中无法提取可扩增的DNA。总之,这项工作表明,将尼龙棉签与DNA稳定防腐剂一起使用,对在高温下储存长达一个月的样品中可扩增DNA的数量产生了切实的影响,并且结合这些方法具有相当大的法医实用性。
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引用次数: 0
Identifying the end user requirements for a novel, field-based environmental DNA (eDNA) forensic collection method 确定最终用户对一种新颖的、基于现场的环境DNA (eDNA)法医采集方法的需求
IF 2 4区 医学 Q2 MEDICINE, LEGAL Pub Date : 2025-12-03 DOI: 10.1016/j.scijus.2025.101371
Matthew Lewis , Katie Lainé , Kirstie Scott , Bernd Hänfling , Nick Dawnay
Environmental DNA (eDNA) refers to the DNA derived from species and found in environmental samples such as water, air and soil. eDNA has been used in biodiversity and conservation studies but not routinely implemented in forensic investigations. Currently physical surveys are implemented at freshwater pearl mussel, Margaritifera margaritifera, crime scenes which are costly, invasive, reliant on practitioner experience and susceptible to false negative results. The aim of this study was to identify key end-user requirements for a novel eDNA sampling method collected through stakeholder discussion and questionnaire. Stakeholder discussions identified the following key criteria for method development: the end-user group, ease of collection, risk to end-user, speed of collection, cost of collection, and data accuracy. These criteria were used to develop three possible eDNA collection approaches: basic (2L water bottles for bankside collection), intermediate (Whirl-Pak® bags for bankside collection), and advanced (onsite Sterivex™ filtration equipment to use on samples collected by entering the watercourse). To compare the three possible methods in respect to their ease and risk, a questionnaire was disseminated to government officers, police officers, postgraduate students and university staff with expertise in either wildlife crime and/or eDNA approaches (N = 63). The time taken to collect samples together with the associated costs of equipment and analysis time were also calculated. Finally, impact of collection method on data accuracy was assessed through the development of a qPCR assay to measure the sensitivity, specificity and concentrations across two different DNA fragment sizes from hatchery reared populations of M. margaritifera. End-user questionnaire data reveals a preference for the basic and intermediate methods in respect to their ease of use and identified risks. Furthermore, these sampling strategies were quicker and cheaper to use although the advanced method could offer logistical benefits in terms of longer preservation times and reduced laboratory processing time. In respect to impact on data accuracy, the advanced method also showed poor detection rates of eDNA, a surprising result given the popularity of the method in conservation studies. Together, these data show the benefits of a co-developed and usable method for police and conservation first responders to use at wildlife crime investigations concerning M. margaritifera habitat destruction. Further research to develop a robust qPCR assay and assess the usability of the basic and intermediate collection methods on wild populations is recommended to determine the overall effectiveness of the described methods.
环境DNA (Environmental DNA, eDNA)是指从物种中提取并在水、空气和土壤等环境样本中发现的DNA。eDNA已用于生物多样性和保护研究,但在法医调查中并不经常使用。目前的物理调查是在淡水珍珠贻贝,Margaritifera Margaritifera,犯罪现场进行的,这些调查费用昂贵,侵入性强,依赖于医生的经验,容易产生假阴性结果。本研究的目的是通过利益相关者讨论和问卷调查,确定一种新型eDNA采样方法的关键终端用户需求。涉众讨论确定了方法开发的以下关键标准:最终用户组、收集的便利性、对最终用户的风险、收集的速度、收集的成本和数据的准确性。这些标准用于开发三种可能的eDNA收集方法:基本(2L水瓶用于河岸收集),中间(Whirl-Pak®袋用于河岸收集)和高级(现场Sterivex™过滤设备用于通过进入水道收集的样品)。为了比较这三种可能的方法的便利性和风险,研究人员向政府官员、警察、研究生和大学工作人员分发了一份问卷,这些人员具有野生动物犯罪和/或eDNA方法的专业知识(N = 63)。还计算了采集样品所需的时间以及相关的设备成本和分析时间。最后,通过开发qPCR方法来评估收集方法对数据准确性的影响,以测量来自孵化场饲养的M. margaritifera种群的两种不同DNA片段大小的敏感性、特异性和浓度。最终用户问卷调查数据显示,就其易用性和可识别的风险而言,他们更倾向于基本和中间方法。此外,尽管先进的方法可以在更长的保存时间和更短的实验室处理时间方面提供后勤方面的好处,但这些采样策略使用起来更快、更便宜。在对数据准确性的影响方面,先进的方法也显示出较低的eDNA检出率,考虑到该方法在保护研究中的普及,这是一个令人惊讶的结果。总之,这些数据显示了一种共同开发和可用的方法对警察和保护第一响应者在野生动物犯罪调查中使用的好处。建议进一步研究开发强大的qPCR检测方法,并评估基本和中间收集方法对野生种群的可用性,以确定所述方法的总体有效性。
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