The Forensic Entomology Case Report-A Global Perspective.

Forensic practitioners analyzing entomological evidence are faced with numerous challenges when presenting their findings to law practitioners, particularly in terms of terminology used to describe insect age, what this means for colonization time of remains, and the limitations to estimates made. Due to varying legal requirements in different countries, there is no standard format for the entomological case report prepared, nor any guidelines as to the sections that are required, optional or unnecessary in a case report. The authors herein propose sections that should be considered when drafting an entomological case report. The criteria under which entomological evidence is analyzed are discussed, as well as the limitations for each criterion. The concept of a global, standardized entomological case report is impossible to achieve due to national legislative differences, but the authors here propose a basic template which can be adapted and changed according to the needs of the practitioner. Furthermore, while the discussion is fairly detailed, capturing all differences between nations could not be accomplished, and those initiating casework for the first time are encouraged to engage other practicing forensic entomologists or professional associations within their own nation or region, to ensure a complete report is generated that meets lab or national requirements, prior to generating a finalized report.

Preliminary data on pupal development, lifespan and fertility of Cynomyamortuorum (L., 1761) in Belgium (Diptera: Calliphoridae).

BACKGROUND: The calliphorid Cynomyamortuorum (L., 1761) is a species of forensic interest, present mainly in the Palaearctic Region. Nearly nothing is known about its life history. NEW INFORMATION: We provide here the first data regarding pupal weight evolution during the pupal stage, female fertility and life expectancy of the species. At 22°C under a variable regime of temperatures, the egg-to-adult development time was an average of 18.05 ± 0.72 and 18.47 ± 0.67 days for females and males, respectively, in the control group. The pupal stage represented 56.7% of the total development. The development time from egg to adult and the duration of the pupal stage were significantly longer for males than for females. The measurement of pupal weight at the start of the pupal period revealed that female pupae were significantly lighter than male pupae by nearly 20%. This difference between the sexes was also observed for the dry weight of adults. An average decrease of 8.75% was observed throughout the first 8 days of the pupal stage, after which most adults started to emerge. The tested females produced an average of 176.13 ± 66.62 eggs throughout the egg-laying period. The average lifespan after emergence was 12.10 ± 4.09 days for females and 12.60 ± 2.95 days for males, with a median of 12.50 days for both sexes.

DNA barcoding and the differentiation between North American and West European Phormia regina (Diptera, Calliphoridae, Chrysomyinae).

Phormia regina (the black fly) is a common Holarctic blow fly species which serves as a primary indicator taxon to estimate minimal post mortem intervals. It is also a major research model in physiological and neurological studies on insect feeding. Previous studies have shown a sequence divergence of up to 4.3% in the mitochondrial COI gene between W European and N American P. regina populations. Here, we DNA barcoded P. regina specimens from six N American and 17 W European populations and confirmed a mean sequence divergence of ca. 4% between the populations of the two continents, while sequence divergence within each continent was a ten-fold lower. Comparable mean mtDNA sequence divergences were observed for COII (3.7%) and cyt b (5.3%), but mean divergence was lower for 16S (0.4-0.6%). Intercontinental divergence at nuclear DNA was very low (≤ 0.1% for both 28S and ITS2), and we did not detect any morphological differentiation between N American and W European specimens. Therefore, we consider the strong differentiation at COI, COII and cyt b as intraspecific mtDNA sequence divergence that should be taken into account when using P. regina in forensic casework or experimental research.

Utility of GenBank and the Barcode of Life Data Systems (BOLD) for the identification of forensically important Diptera from Belgium and France.

Fly larvae living on dead corpses can be used to estimate post-mortem intervals. The identification of these flies is decisive in forensic casework and can be facilitated by using DNA barcodes provided that a representative and comprehensive reference library of DNA barcodes is available. We constructed a local (Belgium and France) reference library of 85 sequences of the COI DNA barcode fragment (mitochondrial cytochrome c oxidase subunit I gene), from 16 fly species of forensic interest (Calliphoridae, Muscidae, Fanniidae). This library was then used to evaluate the ability of two public libraries (GenBank and the Barcode of Life Data Systems - BOLD) to identify specimens from Belgian and French forensic cases. The public libraries indeed allow a correct identification of most specimens. Yet, some of the identifications remain ambiguous and some forensically important fly species are not, or insufficiently, represented in the reference libraries. Several search options offered by GenBank and BOLD can be used to further improve the identifications obtained from both libraries using DNA barcodes.

A fluorescent microscopy-screening test for efficient STR-typing of telogen hair roots.

Nuclear DNA-profiling from human hairs is a well-known technique in forensic investigations, but its success rate is quite low with some hair types. As nuclear DNA (nuDNA) extracted from telogen hair roots is in short supply and is often degraded, a simple and effective method of estimating the number of nuclear DNAs in telogen roots has been developed. DAPI, a fluorescent, non-destructive DNA stain, allows the visualization of "nuclei" (DAPI-positive spots the shape and size of the human follicular cell nuclei) and does not interfere with subsequent PCR analyses. We stained 3242 telogen roots from 27 donors. Surprisingly, of the 2572 club roots without any soft tissue remnants 11% contained visible "nuclei" and 3.3% even contained many. At the same time 57% of the 670 telogen roots with soft tissue remnants did not show any fluorescent "nuclei". We analysed the STR-profile of some of the roots selected by the DAPI screening, i.e. 132 telogen roots without soft tissue remnants, with a success rate of 79%. Our proposed screening method allows the DNA laboratory to analyse nuclear DNA only in the most promising hair roots.