Alteration of expirated bloodstain patterns by Calliphora vicina and Lucilia sericata (Diptera: Calliphoridae) through ingestion and deposition of artifacts

Abstract: Bloodstain pattern analysis can provide insight into a sequence of events associated with a violent crime. However, bloodstain pattern analysis can be confounded by the feeding activity of blow flies. We conducted two laboratory experiments to investigate the relationships between Lucilia sericata (green bottle fly) and Calliphora vicina (blue bottle fly), expirated bloodstains, and pooled bloodstains on a range of surfaces (linoleum, wallpaper, textured paint). C. vicina and L. sericata changed bloodstain pattern morphology through feeding and defecation. They also deposited artifacts in rooms where blood was not present originally. Chemical presumptive tests (Hemastix^®, phenolphthalein, leucocrystal violet, fluorescein) were not able to differentiate between insect artifacts and bloodstains. Thus, C. vicina and L. sericata can confound bloodstain pattern analysis, crime scene investigation, and reconstruction. Crime scene investigators should be aware of these fundamental behaviors, and the effects that blow flies can have on expirated and pooled bloodstain patterns.     

Changes in the morphology and presumptive chemistry of impact and pooled bloodstain patterns by Lucilia sericata (Meigen) (Diptera: Calliphoridae)

Bloodstain pattern analysis can be critical to accurate crime scene reconstruction. However, bloodstain patterns can be altered in the presence of insects and can confound crime scene reconstruction. To address this problem, we conducted a series of controlled laboratory experiments to investigate the effect of Lucilia sericata (Meigen) on impact bloodstains and pooled bloodstains in association with three combinations of common surfaces (linoleum/painted drywall, wood floor/wallpaper, and carpet/wood paneling). L. sericata fed from the pooled bloodstains and added insect stains through regurgitation and defecation of consumed blood. L. sericata formed defecatory trails of insect stains that indicated directionality. Defecatory stains fluoresced when viewed at 465 nm with an orange filter. These observations differed from Calliphora vicina insect stains because feeding on blood spatter was not observed and trails of insect stains were formed by L. sericata. The fluorescence of defecatory stains can be used as a method to detect insect stains and discriminate them from real bloodstains.     

A novel method for ABO genotyping using a DNA chip

ABO genotyping is often performed to identify the blood type of decomposed samples, which is difficult to be determined by a serological test. In this study, we developed a simple method for ABO genotyping using a DNA chip. In this method, polymerase chain reaction-amplified and fluorescent-labeled fragments in the ABO gene and primate-specific D17Z1 were hybridized with DNA probes on a chip designed to detect single nucleotide polymorphisms (SNPs) in the ABO gene and part of the D17Z1 sequence. Using blood samples from 42 volunteers and 10 animal species, we investigated whether the chip could be used to detect SNPs in the ABO gene and the D17Z1 sequence. This method was then applied to various forensic samples, and it was confirmed that this method was suitable for the simultaneous analyses of ABO genotyping and species identification. This method fulfills the recent need for the development of rapid and convenient methods for criminal investigations.     

Volume Determination of Fresh and Dried Bloodstains by Means of Optical Coherence Tomography

The volume of bloodstains found on crime scenes may help forensic investigators reconstruct the location and kinematics of bloodletting events, as stain size, volume, and impact velocity are related. Optical coherence tomography was used as a method to determine the volume and volume ratio of dried and fresh bloodstains on both glass and irregular surfaces or deposited with an impact velocity. The volume of blood drops deposited on smooth glass surfaces was measured within a deviation of 2%. This deviation increased for droplets on irregular surfaces or deposited with an impact velocity. The volume ratio of dried and fresh bloodstains was equal to 19–28% depending on the individual donor and on the use of an anticoagulant. Optical coherence tomography is a good method to determine the volume of fresh and dried bloodstains in laboratory conditions and allows accurate determination of the dry/fresh ratio.     

紫外可见漫反射光谱法无损确证血痕的研究

目的建立一种快速确定血痕的方法。方法将人血、猪血、鸡血、羊血等制成样品纱布,利用紫外可见分光光度计扫描反射光谱进行检测,同时考察了人血的最低检出量。结果血痕的种属与浓度不影响紫外可见反射光谱波峰波谷位置;人血的最低检出量为1.0μL。结论该方法确证血痕具有快速、灵敏、准确、不损坏检材的特点。     

Quantification of psilocin in human whole blood using liquid chromatography–tandem mass spectrometry (LC–MS/MS)

There has been burgeoning interest in psilocybin-use for the treatment of various neurological and neurodegenerative diseases. Psilocybin is mistakenly perceived as the principal pharmacologically active compound due to its high concentrations found in magic mushrooms; however, it is the prodrug of psilocin. Despite the expanding body of clinical research seeking to understand the pharmacodynamic/pharmacokinetic properties of psilocin, and its role in inducing dramatic changes to cognitive function, there has not been a corresponding increase in the development of sensitive analytical methods that can quantify psilocin in different biological fluids. Existing analytical methods have been developed using plasma, serum, and urine as the matrix of choice, but with the unknown blood-to-plasma ratio of psilocin, any pharmacokinetic conclusions drawn solely on plasma data may be misleading. Thus, the main objective of this study is to develop the first analytical method that utilizes SPE and LC–MS/MS to quantify psilocin in human whole blood. The SPE procedure yielded a high recovery efficiency (≥89%) with minimal matrix effects. The method was validated according to ANSI/ASB 036 guidelines. Linearity was between 0.7–200 ng/mL and encompassed previously reported ranges found in plasma/serum. Bias, within- and between-run precision for all quality controls met ANSI/ASB 036 acceptability criteria. Endogenous/exogenous interferences and carryover were negligible. Psilocin stability was assessed at 4°C over 48 h and was considered stable. Although a proof-of-concept study will need to be performed to characterize the method, this analytical workflow was able to detect and quantify psilocin in human whole blood at low limits of quantification.     

Assessment of body fluid identification and DNA profiling after exposure to tropical weather conditions

Despite current advances in body fluid identification, there are few studies evaluating the effect of environmental conditions. The present work assessed the detection of body fluids, blood, semen, and saliva, through lateral flow immunochromatographic (LFI) tests, exposed to tropical weather conditions over time, also evaluating the possibility of obtaining STR (short tandem repeat) profiles and identifying mitochondrial DNA (mtDNA) polymorphisms. Blood, semen, saliva samples, and mixtures of these fluids were deposited on polyester clothes and exposed to open-air tropical weather conditions for 1 month. The test versions from LFI (SERATEC®, Germany) Lab and crime scene (CS) used for the detection – one per each body fluid type – demonstrated that it is possible to identify body fluids and their mixtures up to 14 days after deposition. At 30 days, blood and semen were detected but not saliva. Full STR profiles were obtained from 14-day-old blood samples, and partial profiles were obtained from the remaining samples. It was possible to sequence mtDNA in the samples previously analyzed for STR profiling, and haplogroups could be assigned. In conclusion, this study demonstrated for the first time the possibility of body fluid identification and DNA profiling after exposure to tropical weather conditions for 1 month and also demonstrated the value of mtDNA analysis for compromised biological evidence.     

动脉喷溅血迹动力学及形态学的模拟实验观察

目的观察不同压力、管径对在水平方向上形成的喷溅状血迹的喷射距离及形态的影响。方法建立室内现场环境,利用自制的模拟喷射装置,在60～210mmHg范围内设6种压力,在1.5～4.5mm范围内设4种管径条件,于50、100、150cm 3种高度,制作动脉喷溅血迹。观察水平面上不同喷射高度的喷溅血迹的分布范围、血迹大小及形态特征,并测量其距离,采用SPSS 13.0统计软件进分析,并进行方差检验。结果在不同高度,血液喷射距离随压力增大而增加,随管径增大而减小,两因素间存在交互作用,距离与压力及管径存在较好的线性关系,可得出回归方程。而喷溅血迹的形态特征总体趋势为:高度越高圆形血滴直径约大;管径越粗,边缘毛刺状突起越明显;压力越大,喷射距离越远,且卫星状血迹逐渐增多,拖尾更拉长;随管径的增加,出现椭圆形血滴所需的压力越高。结论动脉喷溅血迹的形态及喷射距离的观察与测量,可用于推断命案现场喷溅源位置及人体受损伤时的体位。     

Method Development and Validation for Determining 1,3-Bu- tadiene in Human Blood by Gas Chromatography-Mass Spec- trometry and Head-Space Gas Chromatography

To develop a simple,validated method for identifying and quantifying 1,3-butadiene(BD) in human blood by gas chromatography-mass spectrometry(GC-MS) and head-space gas chromatography(HS-GC).BD was identified by GC-MS and HS-GC,and quantified by HS-GC.The method showed that BD had a good linearity from 50 to 500μg/mL(r0.99).The limits of detection and quantification were 10 μg/mL and 50 μg/mL,respectively.Both the intra-day precision and inter-day precision were 6.08%,and the accuracy was 96.98%-103.81%.The method was applied to an actual case,and the concentration of BD in the case was 242 μg/mL in human blood.This simple method is found to be useful for the routine forensic analysis of acute exposure to BD.     

超高效液相色谱串联质谱法检测血液中的林可霉素

目的建立超高效液相色谱串联质谱法(UPLC-MS/MS)测定全血中林可霉素的方法。方法样品血使用3mL水提取,涡旋离心,上清液过HLB柱,用10%甲醇溶液淋洗,甲醇洗脱,洗脱液用于UPLC-MS/MS分析。采用ACQUITY UPLC○R HSS T3色谱柱分离,采用电喷雾多反应监测模式(MRM)检测。结果以林可霉素母离子406.987(m/z)和子离子126.011及359.009(m/z)定性、定量。加标回收率在104.98%～120.74%。在S/N≥3的情况下,最低检出限为55.4pg/mL。结论本方法分析速度快,灵敏度高,准确度高,重现性好,可在法庭科学应用推广。