ArticlePDF Available

Isolation and characterization of human DNA from bed bug, Cimex lectularius L.,(Hemiptera: Cimicidae) blood meals

Authors:

Abstract

The ability to identify individual human hosts based on analyses of blood recovered from hematophagous insects is beneficial for both medical and forensic entomology. Bed bugs, Cimex lectularius L. (Heteroptera: Cimicidae), may have several advantages over other blood-feeding arthropods for forensics because they do not remain on the host after their blood-feeding activity and remain in close proximity to a crime scene. Successful isolation, amplification, and sequencing of human DNA obtained from adult bed bugs is reported for the first time from this study. Engorged bed bugs were recovered from a human volunteer and from field collected samples from New York, New York, and Brazos County, Texas. Samples were preserved by drying, stored in 70% ethanol, or freezing at -20°C. DNA was extracted from individual insects, and polymerase chain reaction was conducted using short tandem repeat (STR), human mitochondrial DNA (mtDNA) hypervariable region (HVR1), and insect mtDNA 16S markers. Amplification of a STR marker used in forensic investigations, D18S51, a HVR1 marker, and an insect mtDNA 16S marker was successful. These results demonstrate that DNA isolated from bed bugs is qualitatively and quantitatively sufficient for DNA typing and could be helpful to identify individuals for forensic analysis.
A preview of the PDF is not available
... Human DNA has also been isolated from common bed bugs (Cimex lectularius) after feeding, and the concentration of extracted DNA was adequate for DNA profiling [9,10]. Moreover, a complete human DNA profile has been generated from a DNA extract recovered up to 72 h after these bugs' post-blood meal [11]. ...
... Since this study's main aim was merely to detect human DNA traces within tropical bed bugs, only one common STR locus -the D18S51 marker (selected as it is easy to use in amplification), and one standard mitochondrial marker -HVR1, were used. The selection of markers was also adapted from the study by Szalanski et al. on the isolation and characterization of human DNA from common bed bugs (Cimex lectularius L.) [10]. ...
... PCR cycles for both HVR1 and D18S51 markers were adapted from Szalanski et al. [10]. For the HVR1 marker, PCR cycles consisted of 28 cycles of denaturing at 94ºC for 30 s, annealing at 60ºC for 30 s, an extension at 72ºC for 45 ssconds, and a final extension at 60ºC for 30 min. ...
Article
Full-text available
The ability to isolate and generate a DNA profle from human DNA recovered from tropical bed bugs (Cimex hemipterus) for identifying individuals can be useful for public health, forensic, and medical entomology. In this study, genomic DNA was recovered from both male and female bed bugs at every time interval tested (0, 1, 3, 5, 7, 14, 30, and 45 days post blood meal). The total DNA concentrations recovered from male bed bugs ranged from 12.93 to 65.97 ng/µL, while the total DNA concentrations from female bed bugs ranged from 8.93 to 44.53 ng/µL. However, based on the results from the BLAST search and PCR products, human DNA could be detected from female bed bugs at 0, 3, 5, 14, and 30 days post blood meal using the D18S51 marker. Concentrations of PCR products of the D18S51 locus from male bed bugs ranged from 4.20 to 35.50 ng/µL, whereas, for female bed bugs, concentrations ranged from 4.31 to 22.47 ng/µL. These were generally higher compared to the PCR products of the frst hypervariable part (HVR1) marker. The results indicate the HVR1 locus was less sensitive than the D18S51 locus
... However, human DNA isolated from an insect would have to be (1) stable and intact long enough to be useful in a forensic investigation and (2) unambiguously identifiable to an individual host. Researchers have successfully isolated, amplified, and profiled human mitochondrial DNA (mtDNA) from blood-feeding insects, including bed bugs [5] and human crab louse (Pthirus pubis) [6], and from maggots of the shiny blue bottle fly (Cynomyopsis [=Cynomya] cadaverina) feeding on human tissues [7,8]. Additional research has been conducted to isolate and profile human nuclear DNA from the blood meals of other insects such as human lice (Pediculus humanus capitis) [9] and mosquitoes (Culicidae) [10], and from maggots [11][12][13][14]. ...
... Szalanski et al. [5] demonstrated that DNA could be isolated from recently blood-fed bed bugs and it is qualitatively sufficient for DNA genotyping. However, to date, there are no documented reports about successful human blood identification and/or full human STR typing from a bed bug fed on human female, male, or pooled (female:male) blood. ...
... Mites and insects in the subclass of arthropods are commonly used in the fields of forensic sciences to estimate the time of death and place of the corpse [9]. There are several reports that identified the hosts' genetic material by DNA analysis of macro-sized insects that suck blood and feed on tissue [10][11][12][13]. ...
... Mumcuoğlu et al. [11] obtained the DNA profiles of the hosts by collecting the head and body of lice. Szalanski et al. [13] detected human DNA in bloodsucking bedbugs collected from the beds and the surroundings. ...
Article
Full-text available
House dust mites are microscopic arthropods commonly found in many households. Since they feed on the flakes of shed human skin, human genetic material is expected to be present in these creatures. We conducted a study to find out if house dust mites can carry the DNA of the house occupants. If this were true, could human DNA isolated from the mites, obtained from a crime scene, be used as evidence in court? Dust samples were collected from 27 dwellings using a modified vacuum cleaner. Mites have been isolated from these dust samples by a floating method. Blood samples were obtained from 40 individuals residing in these households. DNA was isolated both from the blood samples and collected mites, and then amplified to detect 9 miniSTR loci by PCR using AmpF&TR MiniFiler (Applied Biosystems) kit. PCR products were processesed in capillary electrophoresis to obtain electrophoregrams. In this study, we detected human DNA in 10.25% of house dust mites collected from 26 out of 27 dwellings (96.3%). Total of 1740 mite samples including different mite species were obtained from the dust samples. DNA profiles of 4 individuals among 39 people (10.25%) from these 26 homes showed an exact match with those found in the mite samples from the same house. Minimum 2 miniSTR loci and maximum 8 mini-STR loci were genotyped from the collected samples in 21 out of 26 houses aforementioned. We identified human DNA in house dust mites suggesting that one can investigate a crime by analyzing DNA samples from house dust mites found in a crime scene, and comparing them with the DNA profiles obtained from victims and suspects. Therefore, house dust mites can be used as evidence in fields of forensic sciences.
... However, in 2018, a new potential use for Cimex detection dogs became apparent as bed bugs proved to be viable forensic physical evidence by yielding human DNA quantitation to help establish associations between perpetrators/victims to the scene of a crime [32]. Previous work had established that it was possible to extract human DNA from blood-sucking bed bugs [33]. ...
... A phylogenetic investigation based on the sequencing of mitochondrial genes is restricted to related species due to its high nucleotide substitution (Boore & Brown, 1998;Rawlings et al., 2001). In the present phylogeny, the partial mt 16S rRNA gene is used to strongly support the interrelationship between the recovered species with other hemipteran taxa at the generic level, this agreed with Hypša et al. (2002), Ribera et al. (2003), and Szalanski et al. (2006) Our findings confirmed that the order Hemiptera clustered four suborders, consistent with previous studies by Gordon et al. (2016) and Walker et al. (2016), who reported three lineages of herbivorous taxa of Sternorrhyncha, Auchenorrhyncha, and Coleorrhyncha were clustered by the species-rich insect order Hemiptera. ...
Article
Full-text available
Insecta is known to be the most diverse group of species, exhibiting numerous forms of endosymbiotic associations. Molecular techniques have provided significant indicators for insect–microbe interactions. The present study aimed to register one of the true bugs of pentatomomorpha and clarify its taxonomic position through phylogenetic analysis of the partial 16S rRNA gene region. A maximum likelihood analysis retrieved a generally well‐supported phylogeny based on Tamura 3‐parameter model. Based on the partial mitochondrial 16S rRNA gene sequences, a phylogenetic study of suborder Heteroptera relationships within Hemipteras' order was constructed. Sequences of 221 bases of the 3ʹ end of the gene from 28 species within 16 families were analyzed. This analysis and bootstrap confidence revealed two major clades comprising four suborders within Hemiptera, with a close relationship between Heteroptera + (Sternorrhyncha + (Auchenorrhycha + Coleorrhyncha)). Infraorder Pentatomomorpha is forming a sister group with a substantial bootstrap value to Cimicomorpha. Pyrrhocoroidea forms a sister relationship with Lygaeoidea + Coreoidea. There is a close relationship between Largidae and Pyrrhocoridae within Pyrrhocoroidea. The results show that the present species is firmly embedded in the genus Arhaphe with 94.35% sequence resemblance to its congeners. Besides, the recovered hemipteran species considered a potential model group for studying different symbionts. We propose both phylogenetic and ecological evolutionary developmental biology viewpoints for a more synthetic understanding of insect populations' molecular evolution. Highlights • • Our data highlighted the need of further taxonomic studies combining both morphological and molecular methods. • • Elevate the higher classification level of various species within different families.
... Research conducted in several countries shows that one in 14 women has been sexually abused and that there is a great under-reporting of this type of crime [7]. Pioneering studies in the isolation and amplification of human DNA using a forensic STR marker (D18S51) were performed on samples gathered from Cimex lectularius L bed bugs [8]. With the use of larvae of Calliphora vicina, which fed on human cadaver, it was possible to recover human autosomal DNA and Y chromosome [9]. ...
Article
The number of sexual crimes in Brazil, as in several other countries, is very high. In many of these crimes the women raped are murdered and their bodies are found days later, in an advanced state of decomposition, with intense cadaverous fauna. Forensic Entomology studies insects and other arthropods that can be used in the expert analysis of various types of crimes. Diptera, the order of insects that comprises the two-winged or true flies, represents one of the largest known groups of insects and is the principal source of cadaveric entomofauna. Members of its Calliphoridae family are observed in cadavers in all phases of decomposition. The retrieval and identification of human Y-STR DNA from the gastrointestinal tract of Calliphoridae species Chrysomya albiceps maggots and pupae can provide a good tool for the gathering of evidence in sexual crime investigations involving rape and death, in which the abandoned victim's body is found in a putrefied state. In this study, the animal model used was a female pig, Sus scrofa, which was sacrificed in a forested area with three shots from a .40 calibre Taurus pistol, and inoculated with semen to its anal and vaginal regions, simulating rape and homicide. During decomposition, 20 to 80 maggots were collected every 24 hours and preserved in 70% alcohol, totalling 289 maggots and 157 pupae (446 immatures) over a period of 14 days (336 hours) of decomposition. Each maggot was then dissected for removal of the digestive tract, which was placed in extraction buffer. The molecular phase proceeded with extraction, quantification, amplification and capillary electrophoresis of samples, testing 16 STR loci of the Y chromosome. It was possible to establish a partial Y-STR DNA profile, with the amplification of up to eight sites, by considering a combination of the samples taken at hours 144 h, 168 h, 192 h, 216 h, 240 h, 288 h, 312 h and 336 h.
... Forensic molecular entomology has proven to be a promising scientific tool; Szalanski et al. (2006) performed the first isolation and amplification of human DNA from the bedbugh Cimer lectularus (Hemiptera, Cimicidae) by utilization of the forensic STR marker D18S51. Similarly, human DNA was recovered from fly larvae that fed on a human corpse, obtaining a complete autosomic and Y-STR profile from various samples (Di Luise et al., 2008). ...
Article
Full-text available
Worldwide, several women become victims of rape every day. Many of those women are also murdered, with their bodies sometimes being found in an advanced state of decomposition, resulting in loss of evidence important to criminal investigations. Diptera is one of the main orders associated with human body decomposition. Fly species that belong to the family Calliphoridae are usually scavengers and are frequently found on decomposing bodies, thereby playing an important role in forensic ntomology. The recovery and genotyping of human Y-STR DNA from the gastrointestinal contents of the calliphorid Chrysomya albiceps larvae has promising applications in the investigation of sexual crimes, such as rape, and in cases of murder and abandonment of the victim’s body, which may be found in a state of decomposition. We studied this species of fly with the aim of supporting such investigations. After establishment of a colony, larvae were fed with decomposing human semen mixed in ground bovine meat (1 mL per 200 g beef). Larvae (10–15) were collected every 24 h and kept in 70% ethanol, to give a total of 96 larvae obtained after eight days of decomposition. The digestive system of each larva was resected. Molecular typing was conducted, which comprised sample extraction, quantification, amplification, and capillary electrophoresis with 16 STR loci from the Y chromosome. We succeeded in establishing a Y-STR DNA profile, with amplification of up to 11 loci, from individual samples, or up to 15 loci, when a combination of samples corresponding to the time-points 48, 72, 120, 144, and 192 h was used.
... The use of insects as forensic evidence is also important in order to detect nosocomial myiasis [12,13] or infection, whereby the insects can act as vectors of diseases, highlighting, sometimes, the state of neglect in which some elderly people live. In particular, entomological evidence can provide space-time information regarding events [14] thereby establishing a correlation between the insect, crime scene and victim [15], helping to identify the human remains [16][17][18][19][20][21] and investigating deaths caused by neglect. In some cases, fly species are difficult to distinguish morphologically, especially at the juvenile stage. ...
Article
Rapid and progressive advances in molecular biology techniques and the advent of Next Generation Sequencing (NGS) have opened new possibilities for analyses also in the identification of entomological matrixes. Insects and other arthropods are widespread in nature and those found at a crime scene can provide a useful contribution to forensic investigations. Entomological evidence is used by experts to define the postmortem interval (PMI), which is essentially based on morphological recognition of the insect and an estimation of its insect life cycle stage. However, molecular genotyping methods can also provide an important support for forensic entomological investigations when the identification of species or human genetic material is required. This case study concerns a collection of insects found in the house of a woman who died from unknown causes. Initially the insects were identified morphologically as belonging to the Pediculidae family, and then, human DNA was extracted and analyzed from their gastrointestinal tract. The application of the latest generation forensic DNA assays, such as the Quantifiler® Trio DNA Quantification Kit and the HID-Ion AmpliSeq™ Identity Panel (Applied Biosystems®), individuated the presence of human DNA in the samples and determined the genetic profile.
Article
Insects have historically been used in criminal investigations to provide information in relation to postmortem intervals (PMIs) but the field of forensic entomology is expanding. It is now recognized that insects can act as vectors of human and mammalian DNA through the consumption of biological material, with extraneous DNA able to be extracted and genotyped from all stages of the life cycle, and insect feces and regurgitant (artifacts). To date, DNA recovered from insects have been used to inform investigations into neglect and homicide, link body parts, and to identify victims of crime. It may also potentially be used to identify assailants, confirm the food source of insects to determine their relevance in PMI calculations, determine if a crime has occurred, identify scenes of crimes, and link people to locations and other individuals. However, insects which have consumed biological material may also transfer DNA by traveling to new areas, or depositing it via their artifacts, either within crime scenes or laboratories, or at locations distant to the food source. This could contaminate forensic evidence, confound investigations, and/or falsely incriminate or exclude an individual. Therefore, it is important that there is increased awareness into both the utility of insects as vectors of forensically relevant DNA, and the potential for contamination. This article is categorized under: • Forensic Biology > Forensic DNA Technologies • Forensic Science in Action/Crime Scene Investigation > Crime Scene Examination • Forensic Biology > Interpretation of Biological Evidence • Forensic Biology > Body Fluid Identification
Research
Full-text available
This represents one of several sections of "A Bibliography Related to Crime Scene Interpretation with Emphases in Geotaphonomic and Forensic Archaeological Field Techniques, Nineteenth Edition" (The complete bibliography is also included at ResearchGate.net.). This is the most recent edition of a bibliography containing resources for multiple areas of crime scene, and particularly outdoor crime scene, investigations. It replaces the prior edition and contains approximately 10,000 additional citations. As an ongoing project, additional references, as encountered, will be added to future editions. The collection and analyses of insects, or invertebrates, from crime scenes is generally well known among homicide investigators and death scene investigators. It has been the compiler’s experience, however, that the actual practice of such collection, outside the presence of a forensic entomologist, is still overlooked or avoided. Often, an attitude prevails that that level of information is not necessary given the investigator’s knowledge of when an abduction took place, or a subject’s confession. In other situations, collections are not made simply because the investigators are not properly equipped with tools, chemicals, and packaging materials to collect and kill samples, or are not sure of what to do with, or how to store, live specimens. Entomological evidence is unique in that it is, in most criminal investigations, the only type of non-human evidence consisting of living, moving species. It is the hope of the compiler that this section will offer some answers toward appropriate collection procedures and equipment which are not expensive, do not involve a lot of time, or the need for additional manpower. The proper collection of entomological samples combined with accurate spatial, temporal, and environmental data, can yield valuable information toward determining postmortem intervals (Taphonomy - Decomposition and Time Since Death). Subject/Witness statements might be supported or disproved. Works such as Catts and Haskell (1990), and Lord and Burger (1983) have become standards in the field of forensic entomological procedures. In recent years compilations such as that by Byrd and Castner (2010) have demonstrated the increasing interest in forensic applications of a science which otherwise serves advancements in health and agriculture. Amendt, et al. (2007) offer "Standards and Guidelines" for this field of study. Entomology, as a means of determining post-mortem interval, continues to be scrutinized. This is not a bad thing. Any validation, clarification, or revocation of a forensic technique benefit crime scene interpretation. Many of the works below include laboratory analyses and research. That research goes beyond addressing the timing of a death or deposition (post-mortem interval) to toxicological determinations, interpretations of death scene versus depositional environments, et cetera. Like virtually every category in this bibliography, the study of entomology is contingent upon so many factors at a scene that an understanding of other disciplines is a neccessity. Obviously, taphonomy and pathology are directly related to insect activity on discovered remains. Environmental characteristics such as soils and plants, as well as body position either by accident or intentional, could influence the impact of insects in peri- and post-mortem activity. Again, the reader is refered to other categories such as Taphonomy, Geoarchaeology and Soil Science, and Criminal and Cultural Behavior. (2552 citations)
Article
Full-text available
Restriction fragment length polymorphisms in polymerase chain reaction amplified fragments (PCR-RFLP) of mitochondrial DNA were used to differentiate species of New World screwworms (Diptera: Calliphoridae). Twenty-seven restriction enzymes were screened on five regions of mtDNA. Eleven restriction fragment length patterns differentiated New World screwworm, Cochliomyia hominivorax (Coquerel), from secondary screwworm, Cochliomyia macellaria (F.). Five restriction fragment length patterns were polymorphic in C.hominivorax while all fragment patterns were fixed in C.macellaria. Diagnostic restriction fragment length patterns were used for species diagnosis, whereas intraspecific variable patterns were used to characterize field samples and laboratory strains. The PCR-RFLP technique is flexible with regard to developmental stage of the sample and method of preservation. We were able to characterize specimens of all life stages from egg to adult including larvae preserved in alcohol and pinned adults. PCR-RFLP is rapid and inexpensive, enabling specimens to be characterized within 24 h for less than $2.50.
Article
Full-text available
Because of morphological ambiguity, traditional identification of Reticulitermes Holmgren termites has always been difficult and unreliable. A molecular diagnostic method is presented for differentiating Reticulitermes species occurring in the south central United States, which are economically important urban pests. A 379-bp region of the mtDNA COII gene and a 415-bp region of the mtDNA 16S rRNA gene were amplified using polymerase chain reaction (PCR) and sequenced from Reticulitermes flavipes (Kollar), Reticulitermes virginicus (Banks), Reticulitermes hageni Banks, and Reticulitermes tibialis (Banks). Applying DNA sequence data, the PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of two restriction enzymes each for the COII amplicon and the 16S amplicon, were diagnostic for all of the Reticulitermes species analyzed. Based on putative mutation rates, >87% and 97% of the samples should be successfully identified to species with PCR-RFLP of COII and 16S, respectively. To verify the accuracy of our predictions, we examined unclassified Reticultermes populations from Arkansas, Louisiana, Missouri, Oklahoma, Texas, and Virginia using PCR-RFLP. Applying PCR-RFLP, 97 samples were correctly classified to species. This technique allows the use of field-collected specimens preserved in alcohol and can identify termite specimens regardless of caste. PCR-RFLP, resolved with agarose or polyacrylamide gel electrophoresis, provided an efficient method for identification of Reticulitermes species from the south central United States for diagnostic purposes.
Article
Insect mitochondrial genome (mtDNA) analysis is a powerful tool for the study of population genetics and phylogenetics. In the past few years primer sequences for the PCR amplification of various insect mtDNA genes have been published. The objectives of this study were (1) present new primer sequences for six insect mitochondrial genes and (2) test primers designed in our laboratory and some previously published primers on a wide range of insects to determine if amplification of the target fragment could be obtained. The primers for the amplification of the two ribosomal RNA gene (16S and 12S rRNA) fragments are universal for insects and related groups; the primers for NADH5 and NADH4 dehydrogenase gene fragments and cytochrome c oxidase I gene fragment are applicable broadly.
Article
This paper reports the sequences of eleven D8S1179 and twenty one D18S51 alleles. The D8S1179 alleles ranged in size from 162 bp to 202 bp and increased in size by regular 4 bp increments. They were shown to possess a compound repeat region composed of the tetranucleotides TCTA and TCTG. Alleles at the D18S51 locus ranged in size from 271 bp to 343 bp and possessed a simple repeat region composed of the tetranucleotide AGAA. The majority of alleles increased in size by 4 bp increments corresponding to the addition of one tetranucleotide repeat unit. However, three alleles differed in size by 2 bp from the 4 bp increment as a result of a dinucleotide insertion within the 3' flanking region. These alleles also exhibited an altered 3' flanking sequence in the first four nucleotides following the repeat region. The allelic designations proposed for these loci on the basis of this sequence data are currently being used in a multiplex PCR profiling system employed in a National DNA database in the United Kingdom.
Article
The ability to identify individual human hosts based on analyses of blood recovered from the digestive tract of hematophagous arthropods has been a long-term pursuit in both medical and forensic entomology. Blood meal individualization techniques can bring important advancements to studies of vector-borne disease epidemiology. Forensically, these analyses may aid in assailant identification in violent crime cases where blood-feeding insects or their excreta are recovered from victims or at crime scenes. Successful isolation, amplification, and sequencing of human mitochondrial DNA obtained from adult human crab lice fed on human volunteers are reported. Adult lice were removed from recruited volunteers frequenting inner city health clinics. Live lice were killed by freezing and subsequently air dried at ambient temperature. A saliva sample was obtained from each volunteer and served as a DNA reference sample. Volunteers were afforded free, approved pediculosis treatment. Individual lice were subsequently processed using procedures developed for the extraction of mitochondrial DNA from human hair, teeth, and bone. The resulting DNA was amplified by the polymerase chain reaction and sequenced. Our results point to valuable avenues for future entomological research.
Article
Human DNA was prepared from mosquitoes (Culicidae) which were collected in a room shared by four human individuals. Several insects did not contain human blood and DNA preparation from them was not successful. However, high molecular weight human genomic DNA could be isolated from four insects. HLA-DQalpha and D1S80 analysis showed that the blood from one insect was a mixture from two persons, whereas the others contained blood from single individuals. Human DNA isolated 26 h after ingestion was still suitable for typing. These results showed that DNA isolated from mosquitoes is qualitatively and quantitatively sufficient for DNA typing and could be helpful to identify individuals involved in certain cases of body violence or captivity.
Article
The isolation, amplification, and characterization of human DNA from hematophagous (blood feeding) and necrophagous (carrion feeding) arthropods have been advanced significantly by the development of polymerase chain reaction (PCR) DNA sequencing methodologies. Historically, DNA technology has been successfully utilized to identify individual hosts upon which species of hematophagous arthropods have fed. The analysis of hematophagous insects' gut content blood meals has led to major advances in medical entomology and vector-borne disease epidemiology. In the forensic arena, the ability to apply similar techniques to insects recovered from badly decomposed remains has been greatly enhanced through the advent of mitochondrial DNA (mtDNA) techniques. Mitochondrial DNA analyses have been utilized to identify both the human remains upon which fly larvae (maggots) have fed and the species of the larvae themselves. The preliminary work detailed here demonstrates, for the first time, the successful application of mtDNA sequencing techniques to the analysis of necrophagous beetle larvae. A small sample of sap beetle larvae, Omosita spp. (Coleoptera: Nitidulidae), was collected from human skeletal remains during anthropological examination and analyzed for human DNA using mtDNA sequencing. The beetle larvae yielded mtDNA matching that of the host human bone. The results detailed here further demonstrate the robust nature of human mtDNA and the ability to recover valuable mtDNA evidence from forensically important, late decompositional stage insect species.
Article
In homicides with entomological evidence, it may be important to prove the presumed association of fly larvae to a corpse, especially if it is in doubt whether all maggots used for entomological expertise developed and fed on it. The present study demonstrates for the first time the possibility of analyzing human microsatellite DNA present in the digestive tract of necrophagous larvae that fed on decomposed bodies with a postmortem interval up to four months. The obtained human STR profiles support the association of a maggot to a specific corpse. In addition, the identification of the host species (e.g., animal source like pig) can be achieved by analysis of the cytochrome b gene. Maggots were collected from 13 corpses after various postmortem intervals and STR typing and HVR amplifications were performed using their crop contents. In seven cases, a complete STR profile was established, in two cases, an incomplete set of alleles was obtained, and in four cases, STR typing was not successful. HVR analysis was successful in all cases except one. The time of storage of the maggots and the length of the postmortem interval up to 16 weeks appeared to have no particular influence on the quality of the results.