Forensic entomological examination without a corpse

Source: Rom J Leg Med (2025) 33: 210–215; DOI: 10.4323/rjlm.2025.210

By Kristina Baumjohann, Mark Benecke

.pdf of this article

Abstract: In an apartment, the previous tenant had died. His decomposition fluid had seeped into the wooden floorboards of the old building over a long period of time. The next tenant sent us samples of the wood and various insects that had come from the apartments’ floor. There was a noticeable decomposition odor in the apartment. The landlord wanted to immediately sublet the apartment and denied both the presence of the decomposition smell and a connection between this and the presence of the insects. We had to clarify whether the discoloration in the wooden floor was caused by seepage of corpse fluids and whether the appearance of the insects was related to the deceased previous tenant. The species composition of the insects and an ammonia test was indicative of the presence of at least decomposition fluid which may have come from a corpse.

Keywords: forensic entomology, forensic biology, stains, insects, decomposition.

CASE DESCRIPTION

The client had rented an apartment in which the previous tenant had died and been lying dead on the wooden floor in the apartment for a long time. The wooden floorboards there were discolored. There was a noticeable corpse odor in the apartment. Our client repeatedly found various insects in the now empty apartment. The landlord did not want to commission any extensive renovation work, as is usual in the city in question, owing to the housing shortage – he wanted to rent the apartment out again immediately. He stated that: - There was no corpse odor; only a smell of sanded wood. - The visible stains could have been caused by any liquid, e.g. coffee. - The flies found in the apartment had come from outside by chance, had not developed in the apartment and were not attracted by the decomposition smell of the deceased. Insect and wood samples were obtained to clarify whether the discoloration in the wooden floor was caused by corpse fluids seeping into it and whether the occurrence of the insects was related to the deceased previous tenant. We were able to establish a connection between the appearance of the insects and the corpse fluid that leaked and seeped into the wooden floor (and far below).

MATERIAL AND METHODS

Insects

Insects were examined with a stereomicroscope binocular (Leica MZ12.5, max. 100x) and identification keys [1-5].

Floorboards/odor

Due to the time pressure and suffering of those affected, it was not possible for us to inspect and take samples on site. However, we had photos and a video of the apartment in very good quality. The samples of the wooden floorboards were each packed airtight in three-layer Ziploc bags, with another small airtight bag with wooden material inside.

Each sample of those samples was sent to us in a glass jar (Fig. 1). Some of the wood material had a distinctly dark, charred-looking discoloration (Fig. 2). The wood samples consisted of smaller and slightly larger pieces of wood (Fig. 3).

Ammonia test

A rapid ammonia test (Bosike/Like Sun Company, Essen, Germany) was conducted to exclude coffee and as an indication of putrification liquid [6, 7]. Therefore, a small piece of a darkly discolored area of the wooden floor was swirled in 3 ml of sterile water (disposable vial, AddiPak Company), and put in a sterile, DNA-free centrifuge tube (Greiner Company) for a few minutes (Fig. 4). The ammonia test strip was dipped into it.

RESULTS

Insects

The samples contained the following insect groups and species: - Flesh flies of the genus Sarcophaga spec. (Fig. 5); - Blow fly of the species Calliphora vicina (Fig. 6); - Carpet beetles (skin beetles), probably of the genera Attagenus spec. and Anthrenus spec. (Dermestidae) (Fig. 7); - Larder beetle larvae (Dermestidae) (Fig. 8); - Chalcid wasps, probably of the family Torymidae (Chalcidoidea) (Fig. 9).

Flesh flies (n = 56) were most prevalent in the samples, followed by carpet beetles (n = 41) and chalcid wasps (n = 9), while only one blow fly was present (Table 1). The skin beetle larvae were numerous and we did not count them. Apart from the chalcid wasps, all insects found in the samples are associated with decomposition processes of human and animal bodies [8-11].

While blow flies visit dead bodies in earlier stages of decomposition [12, 13], flesh flies colonize corpses during the entire decomposition process [14-22]. In large cities, this is particularly true for corpses found in apartments.

Carpet and/or skin beetles and their larvae occur in later stages of decay (desiccation) and prefer drying organic tissue (including mummies, stuffed animals, etc.) on which they feed [23-26]. Chalcid wasps are predators of other insects that colonize corpses, such as the blow flies and flesh flies found in the samples [27]. They only appear later during the course of corpse colonization, as they do not colonize the corpse as such, but animals living on the corpse.

Individual adult, dead flies showed holes and erosions, which may have been caused by skin beetles and chalcid wasps. This is a further indication of prolonged colonization and/or the prolonged presence of insects colonizing corpses and attracted by the odor of corpses (Fig. 10).

Odor

As soon as the glass lids (i.e. the outermost packaging of the multi-packaged samples) were opened, the laboratory room was immediately filled with the typical smell of decomposition. All samples already gave off a distinct putrefactive smell through the closed bags. The smell of coffee or similar odors could not be detected in any of the two samples – even when the bags with the wooden samples were opened and the odor was tested directly with the nose.

No chemical analysis were performed as this had neither been asked by the client nor had this been necessary for our investigation.

Ammonia

The rapid ammonia test as an indicator putrefactive liquid gave a clear result after just a few seconds of immersion; according to the manufacturer, this normally takes longer (Fig. 11).

Regarding the landlord’s statements

Corpse odor and “stains”

According to the client, “all employees... [of the landlord] who were in the apartment as well as the... [the landlord’s] crime scene cleaner and pest controller [...] stated that there was no corpse odor, only the smell of sanded wood.” This is not correct for the discolored wood: It smelled very distinctly of a decomposing corpse. Irrespective of this, there may have been a strong odor of sanded wood or another odor in the apartment. However, the odor of the corpse must have been perceptible in the immediate vicinity of the wood.

Flies

The clients state: “On his second visit to the apartment, the crime scene cleaner and pest controller took about 3 - 5 specimens of the flies from the large room to have them identified by a biologist friend. We were not informed of the results of the investigation.” The landlord is said to have stated: “An expert confirmed to us that these flies must have come from outside and could not have developed in the apartment.” This statement is partly correct: The insects may have flown into the apartment from outside because they were attracted by the corpse odor serving as an attractant for these insects. They do not necessarily have to have developed there. They can also “only” feed or lay eggs on putrefying tissue. The presence of the skin beetles and adult flies indicates that the colonization/ flying of insects into the apartment, which was either littered with animal meat or had a human corpse, had been taking place for some time: These are so-called late corpse colonizers. All of the insects we examined only occur at corpse sites with decomposing corpses or in environments with a high meat content in old waste (animal or human corpse parts).

DISCUSSION

The insects studied here are either corpse colonizers or are clearly associated with decomposition processes or animals attracted by them.

We are not aware of any odor that is similar in composition to the type of corpse odor present here. A similar odor can occasionally arise from mountains of garbage, which must then consist mainly of meat scraps. Only then does this odor develop, and then only after sufficiently long “maturing”. The time required for this depends on the temperature: The warmer, the faster. Since the apartment in the video sent to us does not appear to be full of garbage, but empty, there would have to have been a large quantity of garbage with a high meat content.

The ammonia test is triggered by the presence of decomposable substances, not by “normal” wood floors or pieces. This shows that decomposition fluid (and not coffee) must have penetrated the floorboards.

We have a video from the apartment which shows that there are cracks in the floor, including where the body is supposed to have been lying. Decomposition fluid and pieces of tissue from the corpse may have entered the area under the floorboards through these cracks (puddle of fluid). Cadaveric fluid can penetrate the actual wood of unsealed floors and then remain there or under the wooden floorboards owing to the adhesive properties of the semi-liquefied cadaveric material.

From time to time, these kinds of contaminated floors are sealed to “encapsulate” the odor or treated with chemicals to dissolve or mask the odor (for example with the raspberry odor of Maskomal or similar). However, it makes more sense to remove the source of the odor; this is comparable to used baby diapers, which are also thrown away or cleaned and not covered up in order to mask the odor.

The collective occurrence of the insects from the apartment samples certainly indicates the presence of at least decomposition fluid, the odor of which attracted the animals. The putrefactive liquid must have been in the wooden floor in sufficient quantity and presumably over a longer period of time, otherwise only blow flies would be found, which colonize corpses at an early stage. Skin beetles and chalcid wasps appear as later corpse colonizers [28]: Carpet beetles and their larvae use dried organic tissue and dead insects (such as flies) as a food source. However, they are less commonly found in apartments with corpses and piles of garbage, as the soiled areas are usually cleared and cleaned quickly. Chalcid wasps in particular are then naturally no longer attracted: They colonize the juvenile stages of other insects.

The absence of blow fly larvae and their pupae in the samples and on the photos indicate that there is now no fresh, new organic material available for the blow fly larvae to feed on. None was found anyway.

As blow fly larvae usually seek out dark and narrow “hiding places” to pupate, fly pupae and larvae could also be found under floorboards and behind carpet rails. This can occur from time to time and sometimes leads to strange occurrences of flies for the residents. For example, an apartment can be cleared and cleaned, but the pupae of the flies are still hidden in cracks and the flies hatch from the pupae later (see case 2 „Empty Apartment“ [29]).

The “mixture” of dead flies, beetles and chalcid wasps in the apartment concerned here means that the animals were attracted by the smell of corpses and died in the apartment or that there was sufficient food (including corpse fluid). It is irrelevant whether these flies developed in the apartment (from larvae) or whether adult insects have flown into the home. The main reason for their appearance is that they are attracted by the smell of decay.

Conflict of interest

The authors declare that they have no conflict of interest.

References

1. Hackston M. Family Dermestidae; 2014, retrieved from https:// quelestcetanimal-lagalerie. com/ wp-content/uploads/2012/11/ Family-Dermestidae-illustrated-key-to British-species.pdf., June 13th 2024.

2. Hackston M.Key to the british families of superfamily Chalcidoidea (Hymenoptera); 2018, retrieved from https://sites. google.com/view/mikes-insect-keys/mikes-insect-keys/ keysfor-the-identification-of-british-hymenoptera-bees-and-wasps/ keys-for-the-identificaitonof-british-parasitic-wasps/keys-for-theidentification-of-british-chalcidoidea, June 13th 2024.

3. Hackston M. Key to the british superfamilies of Hymenoptera; 2019, retrieved from https:// sites.google.com/view/mikes-insectkeys/mikes-insect-keys/keys-for-the-identification-ofbritishhymenoptera-bees-and-wasps, June 13th 2024.

4. Hackston M. Key to UK species adapted and updated from van Emden (1954) Handbooks for the identification of british insects Vol X, part 4(a), Diptera Cyclorrhapha Calyptrata (1); 2019, retrieved from https://sites.google.com/view/mikes-insect-keys/ mikes-insect-keys/keys-for-the-identification-of-british-true-fliesdiptera/keys-for-the-identificationof-british-sarcophagidae, June 13th 2024.

5. Szpila K. Key for the identification of third instars of european blowflies (Diptera: Calliphoridae) of forensic importance. In: Amendt J, Campobasso CP, Goff ML, editors. Current concepts in forensic entomology. Heidelberg: Springer; 2010. p. 43-56.

6. Van en Oever R. Post-mortem vitreous ammonium concentrations in estimating the time of death. Z Rechtsmed. 1978;80:259-263.

7. Patel D, Dargan R, Burr WS, Daoust B, Forbes S. Identifying the early post-mortem VOC profile from cadavers in a morgue environment using comprehensive two-dimensional gas chromatography. Separations 2023;10:566.

8. Farrell JF, Whittington AE, Zalucki MP. A review of necrophagous insects colonising human and animal cadavers in south-east Queensland, Australia. Forensic Sci Int. 2015;257:149-154.

9. Ren LP, Deng HX, Dong SZ, Li JB, Hu XH, Cai FJ, Guo YD. Survey of indoor sarcosaphagous insect. Trop Biomed. 2017;34(2):284-294.

10. Diaz-Aranda LM, Martin-Vega D, Gomez-Gomez A, Cifiran B, Baz A. Annual variation in decomposition and insect succession at a periurban area of central Iberian Peninsula. JFLM 2018;56:21-31.

11. Al-Khalifa MS, Mashaly AM, Al-Qahtni AH. Insect species colonized indoor and outdoor human corpses in Riyadh, Saudi Arabia. Journal of King Saud University. Science 2020; 32:1812-17.

12. Byrd JH, Castner JL. Forensic Entomology: The utility of arthropods in legal investigations. 1st edition. Boca Raton: CRC Press; 2001. p. 45-46.

13. Bugelli V, Forni D, Bassi LA, Di Paolo M, Marra D, Lenzi S, Toni C, Guisiani M, Domenici R, Gherardi M, Vanin S. Forensic entomology and the estimation of the minimum time since death in indoor cases. J Forensic Sci. 2015;60:525-531.

14. Byrd JH, Castner JL. Forensic Entomology: The utility of arthropods in legal investigations. 1st edition. Boca Raton: CRC Press; 2001. p. 50-51.

15. Cherix D, Wyss C, Pape T. Occurrences of flesh flies (Diptera: Sarcophagidae) on human cadavers in Switzerland, and their importance as forensic indicators. Forensic Sci Int. 2012; 220(1- 3):158-163.

16. Bonacci T, Greco S, Cavalcanti B, Brandmayr P, Vercillo V. The flesh fly Sarcophaga (Liopygia) crassipalpis Macquart 1839 as an invader of a corpse in Calabria (Southern Italy). J Forensic Sci Crimin. 2014;1:1-5.

17. Pinto Vairo K, da Cruz Canepro MF, Correa RC, Preti D, Moura MO. Can Sarcophagidae (Diptera) be the most important entomological evidence at a death scene? Microcerella halli as a forensic indicator. Rev Bras Entomol. 2017;61: 275-276.

18. Battan-Horenstein M, Mulieri PR, Gleiser RM. Evaluation of environmental and local factors on urban sarcosaprophagous Diptera: a case study with Sarcophagidae. Med Vet Entomol. 2020;35(2):158-168.

19. Gunn A. The colonisation of remains by the sarcophagid fly Sarcophaga argyrostoma (Robineau-Desvoidy) (Diptera: Sarcophagidae). Forensic Sci Int. 2020;315:110465.

20. Barbosa TM, Carmo RFR, Mello-Patiu CA, Leal IR, Dias Vasconcelos S. Effects of chronic anthropogenic disturbances on flesh fly (Diptera, Sarcophagidae) assemblages in areas of seasonally dry tropical forest. J Insect Conserv. 2021;25:485-498.

21. Afifi FM, Abdelfattah EA, El-Bossiony GM. Impact of using Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy, 1830) as a toxicological sample in detecting clonazepam for forensic investigation. Egypt J Forensic Sci 2022;12:37.

22. Hediyeloo S, Akbarzadeh K, Rezaei M, Oshaghi MA. Colonization pattern and thermal needs of immature phases of Sarcophaga argyrostoma (Diptera: Sarcophagidae): significance for estimating postmortem interval. Heliyon 2024;10:e26576.

23. Byrd, JH, Castner JL. Forensic Entomology: The utility of arthropods in legal investigations. 1st edition. Boca Raton: CRC Press; 2001. p. 66-67.

24. Charabidze D, Colard T, Vincent B, Pasquerault T, Hedouin V. Involvement of larder beetles (Coleoptera: Dermestidae) on human cadavers: a review of 81 forensic cases. Int Journal Legal Med. 2013;128(6):1021-1030.

25. Gunn A. The exploitation of fresh remains by Dermestes maculatus De Geer (Coleoptera, Dermestidae) and their ability to cause a localized and prolonged increase in temperature above ambient. JFLM 2018;59:20-29.

26. Kadej M, Szleszkowski L, Thannhäuser A, Jurek T. A mummified human corpse and associated insects of forensic importance in indoor conditions. Int J Legal Med. 2020;134:1963-1971.

27. Frederickx C, Dekeirsschieter J, Verheggen FJ, Haubruge E. The community of Hymenoptera parasitizing necrophagous Diptera in an urban biotope. J Insect Sci. 2013;13:32.

28. Benecke M. Six forensic entomology cases: description and commentary. J Forensic Sci. 1998; 43:1303.

29. Benecke M. Six forensic entomology cases: description and commentary. J Forensic Sci. 1998;43:797-805.

Estimation of post mortem intervals in humans

Source: Tagungsband der 12. Wissenschaftlichen Tagung der Gesellschaft für Entwicklungsbiologie e. V. vom 11. - 14. März 1997 in Köln (GER)

A macroscopic application of developmental biology called Forensic Entomology

From Mark Benecke

Klick hier für das .pdf


Insect development is well-studied and thus can be used as a marker for estimation of time intervals. Blowflies and several beetles together with their larvae are used in forensic science to help diagnosing how long a corpse has been exposed to defined environmental conditions.

Most times the forensic entomologist is asked if the insect fauna of an corpse can tell the time and date of a person's dead.

For various reasons, pest control experts as weIl as entomologists and (forensic) medicinal practitioners have studied the correlations between temperature, humidity and speed of development of several insects.

Forensic scientists put together those facts and calculated curves which now allow a precise estimation of the living time of an insect found associated to a corpse. In most cases, length, weight, and macroscopic state (e.g., filling of intestines) of larvae are determined and compared to the standard data curve.

Because of several insects visiting corpses only at a defined state of decomposition, further information about the post mortem interval is given by determination of species. Sometimes it is quite hard to determine species just by their pupae or even worse by their larvae. That is why it can be necessary to breed larvae to adultship. In doing this, developmental biology once again is in focus of research and criminal investigations.

Only a lew laboratories in the world (Rosny sous Bois, Hawaii, Wien, Köln, Quantico among few others) perform(ed) studies concerning the forensic application of insect development. We present our aims to establish a database of developmental processes of insects which are specific for the western German fauna of human corpses, especially for the diptera Lucilia caesar, Calliphora vicina and Calliphora erythrocephala.

Insects, Cadavers and Ink

Quelle: Tattoo Master 01/2014, pages 60-62

By Nacho Brown

The article as .pdf

Mark specialises in forensic entomology; the study of insects and arthropod biology and its application to criminal cases and legal matters - gathering information far criminal cases through the examination of insects extracted from dead bodies. It is dear, that while most people would find removing maggots from decaying bodies a little unsavoury, Mark has a deep respect far the creatures he works with.

"I just love invertebrate animals and I very much love to be in the middle of the circle of life. What I do as a forensic entomologist go es straight to my heart and invigorates all of my senses. Other people may connect with their inner self with high dosages of hallucinogenics, to feel fully embedded in earth and nature, for me my work has the same effect“.

Mark's enthusiasm for his work is lost on many people but he has never let the thoughts of others hold him back. In addition, the simple life free of the clutter and clamour for possessions makes sense to him.

"I never cared. I just do my own thing. I worked for awhile in New York and while there I sometimes lived on food that was left outside hotel rooms for the room service to pick up. Still today my friends make fun of me because I never throw away food and even eat food that other people leave. In Manhattan I worked at the Chief Medical Examiner's office as a forensic biologist and I lived in a basement, the cellar of an apartment.

I really did not need much. I do not own a car, or a flat, or anything except my computers, very old books, dissection microscopes and the like. I get along easily and keep everything low maintenance. I find that this has two good side effects: no one can threaten me by taking things away from me and nobody thinks I want his or her job, or power, or money, or what not. So those who want my work, ask for it. Those who do not need it, do not ask. It was like that from the first day onwards. In most countries I have good and stable working relationships in universities, with the police and so on. It's not a struggle, the fact that I live in my lab.“

In recent years forensics and tattooing have begun to cross over with specialists looking at the state of decay of a tattoo to help determine the time of death and using tattoos to help identify bodies. But Mark still feels there is so much more headway to be made.

"More research needs to be carried out in this area. To get an identification process going, sometimes the police enhance tattoos from bodies of the recently deceased, or from the body parts of on es that are decomposed. And sometimes I also I ask around if anybody knows a particular style or motif so we may find out in which region of the world, or in which tattoo parlour, it may have been done.“

Mark's own body is extensively tattooed with a mixture of styles and techniques from rough and ready home done pieces to detailed neo-classical designs. Mark was twenty years old when he got into tattooing and it all started in the city of Cologne, in Germany.

"This is where the cologne perfume comes from and why cologne perfume is called 'cologne’. No kidding! Anyway, back in the eighties, we had one of the first legal German tattoo studios. The window of the shop, the whole area, was rough and basic. You know, just a few pictures of Japanese tattoos etc.

I went in with all the respect a person can have and asked if they could do a lizard on my shoulder. This was not too weird as the owner, a guy by the name of Dieter, did good, bold, old school tattoos. And his wife, Anke, does flowers and tigers etc. Anke said yes and did it and that was how I got started on the path of body modification.

I think tattoos and body modifications are normal for adults. It's your body so do something with it to make your inner image match your outer body image“.

And Mark's body modification goes beyond tattoos ...

„I have magnets in tow fingers and a RFID chip in my arm.“

The magnets make Mark's fingers sensitive to electric currents and magnetic fields giving him, in effect, a sixth sense and an enhanced awareness of his environment. The radio frequency identification chip is a tiny electrical device, not much bigger than a grain of rice, which can be used to store and transfer information such as a person's personal details. Though, in Mark's case, the RFID chip is programmed to one unique id number, so it cannot be reprogrammed.

Mark's passion for tattooing also goes hand in hand with his nomadic journey across the planet. He's worked in America, visited China on numerous occasions and has been across Europe and South America, all the time getting tattooed as he travelled.

„I’ve got tattoos from all around the world. I even became the first male centrefold in the German 'Tätowiermagazin' because my body art is a patch work from across the globe. And like sailors in former times, every tattoo comes from another place.“

Mark is no stranger to being on the other side of the needle either.

"I do fan tattoos once in awhile, usually my 'Vampire Smiley‘. But also little heart anchors and small stuff. Many students want them but also my friends. I only do them black, rough and dirty. I use a go rotary machine as the old coil machine started to kill my ears and nerves“.

In addition to his forensic work and his tattoo collecting, Mark has also worked with controversial artist and anatomist, Gunther von Hagens.

"I worked for Koerperwelten Body World from 1999-2000. Gunther was there the night before it opened and I was one of the few that night to give everything the final touch. Since then I have become very attached to the exhibit and to his factory in Guben on the Polish border. So much so that I give speeches for the public during Koerperwelten exhibitions in Europe.“

So what does the future hold for a man who has carved out a unique niche in the combined worlds of forensics and tattooing?

"I am like a modern sailor and so I have a lot of sailor tattoos. Hold Fast on my hands, Sailing for Truth on my back, a pirate girl on my leg. I just like to sail and see what happens next. I make no plans.“

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