Detection of fly artifacts from four species of necrophagous flies on household materials using immunoassays

Rivers, David; Cavanagh, Gregory; Greisman, Valerie; Brogan, Rebecca S.; Schoeffield, Andrew

doi:10.1007/s00414-019-02159-1

Cited by 4 publications

(7 citation statements)

References 34 publications

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“…In some cases, the origin of the stain can remain undetermined even after a combined macroscopic ultrastructural approach, such as in the case of darkcolored stains with a flat surface that cannot be explored by SEM, like those we observed on polyester, or stains absorbed by the deposition surface, like those we observed on cotton. In these cases, other non-morphological techniques could be used, such as the recently developed immunodetection with polyclonal antiserum [11,12,14,22]. Even if validation studies are needed on other experimental settings, SEM analysis confirmed to be a promising tool for distinctions of FA from true human bloodstains.…”

Section: Discussionmentioning

confidence: 99%

“…However, the main problem associated with visual or ultrastructural stain morphology is that baseline data on fly stains deposited on materials typical of crime scenes, such as fabrics, metal, glass, or plaster is still scarce [11]. Moreover, the morphologic features of FA related to the activity of a fly may differ from the FA produced from other fly species [12].…”

Section: Introductionmentioning

confidence: 99%

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Morphological characterization using scanning electron microscopy of fly artifacts deposited by Calliphora vomitoria (Diptera: Calliphoridae) on household materials

et al. 2021

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Insects found at a crime scene can produce traces referred to as fly artifacts (FA) due to their movement over the corpse and the manner in which they feed upon it. These can be detrimental for carrying out criminal investigations. Confusing a FA with a genuine bloodspot can lead to misinterpretations, also taking into consideration that FA may contain a human DNA profile. The aim of the present study was to employ scanning electron microscopy (SEM) for the analysis of FA produced by Calliphora vomitoria on hard surfaces and fabrics that are commonly present at crime scenes. FA and control bloodstains were produced under experimental conditions on metal, glass, plaster, cotton, and polyester. After macroscopic analysis, FA were examined at standard low (20–40 ×), medium low (300–600 ×), and high ultrastructural (1200 ×) magnification through a SEM Stereoscan 360, Leica, Cambridge. SEM analysis enabled the identification of distinctive features of FA on hard surfaces, namely, amorphous crystals, micro-crystals with a morphology similar to those of uric or micro-crystals with a comparable morphology to cholesterol, absent in controls. Moreover, red blood cells (RBC) were absent in FA but were always present in controls. On cotton, for both FA and controls, the drop was almost completely absorbed and thus indistinguishable from the underlying fabric texture. On polyester, FA showed amorphous/crystal-like deposits and no RBC, as observed on hard surfaces, except for those showing a completely flat surface. SEM analysis appeared to be suitable for differential diagnosis between FA and genuine bloodstains on hard surfaces, although the results may be inconclusive on tested fabrics.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Morphological characterization using scanning electron microscopy of fly artifacts deposited by Calliphora vomitoria (Diptera: Calliphoridae) on household materials

et al. 2021

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“…Since Rivers et al [10][11][12] has reported that only artifacts derived from alimentary canal of the adult flies reacted positively to the immunoassay test, we have chosen to analyze fly artifacts more likely originated from regurgitation or fecal elimination processes, excluding transfer pattern stains produced by tarsi or other body parts, because fly touch DNA was not expected.…”

Section: Discussionmentioning

confidence: 99%

“…In the last years, Rivers et al [10,11] tested a polyclonal antiserum (anti-md3 serum) generated toward a unique cathepsin D proteinase that has been shown to react with regurgitate and defecatory fly artifacts produced by different species of blowflies, but not with transfer patterns or blood controls from humans or other animals. Recently, the study was extended to other stains produced by different species of flies following the consumption of semen, saliva, feces, and urine, showing that more than 94% of fly artifacts reacted positively with anti-md3 [12].…”

Section: Introductionmentioning

confidence: 99%

A DNA-based method for distinction of fly artifacts from human bloodstains

et al. 2021

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Fly artifacts resulting from insect activity could act as confounding factors on a crime scene and interfere with bloodstain pattern analysis interpretation. Several techniques have been proposed to distinguish fly artifacts from human bloodstains based on morphological approach and immunological assay, but a DNA-based method has not been developed so far. Even if in forensic genetic investigations the detection of human DNA is generally the primary goal, fly artifacts can provide useful information on the dynamics of crime events. The present study provides a molecular method to detect fly DNA from artifacts deposited by Calliphora vomitoria after feeding on human blood through the analysis of the mitochondrial cytochrome oxidase gene subunit I (COI). Fly artifacts originated from digestive process and of different morphology spanning from red and brownish/light brown, circular and elliptical stains to artifacts with sperm-like tail or a tear-shaped body were collected. The COI amplification was successfully obtained in 94% of fly artifact samples. The method showed high sensitivity and reproducibility, and no human DNA contamination was observed, offering specificity for use in confirmatory test. This molecular approach permits the distinction of fly artifacts from genuine bloodstains and the identification of fly’s species through the COI region sequencing by protocols usually applied in forensic genetic laboratories.

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“…A sizable amount of work has been done on fly artifacts, including various detection techniques (Rivers et al, 2018; Rivers et al, 2019; Rivers, Cavanagh, Greisman, Brogan, & Schoeffield, 2020) and analyses of their stains (Rivers, Dunphy, Hammerschmidt, & Carrigan, 2020; Rivers & Geiman, 2017; Rivers & McGregor, 2018). Other work relating to entomotoxicology found that both opiates (Introna, Dico, Caplan, & Smialek, 1990) and barbiturates (Levine, Golle, & Smialek, 2000) could be obtained from larvae feeding on remains that had ingested these drugs.…”

Section: Forensic Entomological Literature In the Northeastmentioning

confidence: 99%

A review of Forensic Entomology literature in the Northeastern United States

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Noblesse

et al. 2021

WIREs Forensic Science

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A comprehensive review of forensic entomology literature in the northeastern portion of the United States was conducted searching the earliest possible publication date through early 2020. Thirteen states and the capital were included in this region and yielded 172 publications. As a region, the main areas of the publications were arrival and decomposition, myiasis, and papers yielding information pertaining to biodiversity. However, there was a large fluctuation in total publications by state with Maryland producing over 30 publications, and Delaware and Vermont each producing one. Within this region only 15 publications from seven states were found concerning development, showing a severe deficiency in this category. Additional work is needed in this geographic area of the United States in most categories of work related to forensic entomology research. This article is categorized under: Forensic Biology > Interpretation of Biological Evidence Forensic Anthropology > Time Since Death Estimation Crime Scene Investigation > Crime Scene Examination

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Detection of fly artifacts from four species of necrophagous flies on household materials using immunoassays

Cited by 4 publications

References 34 publications

Morphological characterization using scanning electron microscopy of fly artifacts deposited by Calliphora vomitoria (Diptera: Calliphoridae) on household materials

Morphological characterization using scanning electron microscopy of fly artifacts deposited by Calliphora vomitoria (Diptera: Calliphoridae) on household materials

A DNA-based method for distinction of fly artifacts from human bloodstains

A review of Forensic Entomology literature in the Northeastern United States

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