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Estimating Time of Death with Forensic Entomology

Posted in Forensic Entomology & DNA Barcoding by paibiopai on November 20, 2009

After the initial decay, and the body begins to smell, different types of insects are attracted to the dead body. The insects that usually arrives first is the Diptera, in particular the blow flies or Calliphoridae and the flesh flies or Sarcophagidae.

The females will lay their eggs on the body, especially around the natural orifices such as the nose, eyes(2), and ears(2). If the body has wounds the eggs are also laid in such. Flesh flies do not lay eggs, but deposits larvae instead.

After some short time, depending on species, the egg hatches into a small larvae. This larvae lives on the dead tissue and grows fast. After a little time the larva molts, and reaches the second larval instar. Then it eats very much, and it molts to its third instar. When the larvae is fully grown it becomes restless and begins to wander. It is now in its prepupal stage. The prepupae then molts into a pupae, but keeps the third larval instars skin, which becomes the so-called puparium. Typically it takes between one week and two weeks from the egg to the pupae stage. The exact time depends on the species and the temperature in the surroundings.

The theory behind estimating time of death, or rather the post mortem interval (PMI for short) with the help of insects are very simple: since insects arrive on the body soon after death, estimating the age of the insects will also lead to an estimation of the time of death.


How to estimate age of blowfly eggs, larvae, pupae and adults

When blow flies oviposit, their eggs has come very short in their embryonic development. The eggs are approximate 2 mm in length. During the first eight hours or so there is little signs of development. This changes after that, and one can see the larvae through the chorion of the egg at the end of the egg stage. The egg stage typically lasts a day or so.

The blowfly has three instars of larvae. The first instar is approximately 5 mm long after 1.8 days, the second instar is approximately 10 mm long after 2.5 days, the third instar is approximately 17 mm long after 4-5 days. Identifying the right instar is the easiest part, and is done relatively easy based on size of larvae, the size of the larva’s mouth parts and morphology of the posterior spiracles. The time it takes to reach the different instars depends very much on microclimate, i.e. temperature and humidity.

At the end of the third instar the larva becomes restless and starts to move away from the body. The crop will gradually be emptied for blood, and the fat body will gradually obscure the internal features of the larvae. We say that the larva has become a prepupa. The prepupa is about 12 mm long, and is seen 8-12 days after oviposition.

The prepupa gradually becomes a pupa, which darkens with age. The pupa which are about 9 mm in length are seen 18-24 days after oviposition. The presence of empty puparia should therefore tell the forensic entomologist that the person in question has been dead in more than approximately 20 days. Identification can be done based on the remaining mouth parts of the third instar larvae.

A more precise way to determine age of larvae and eggs is the use of rearing. For example: the body is found with masses of eggs on it, none have hatched. How long time is it since the eggs was oviposited? Note the time of the discovery, note the time when the first 1. instar larvae occur. Subtract the first occurrence time with the discovery time, call this time A. Rear the blow flies to adults, let them mate, let them lay eggs on raw beef liver under conditions similar to the crime scene, take the time from oviposition to the first occurrence of 1. instar larvae. Call this time B. By subtracting B-A, one gets C, which is an estimate of the time since oviposition to discovery. Similar calculations can be done for other instars as well. If one has good base-line data from before under different temperatures and for different species, one only needs to rear the flies to a stage where they can be identified, and that is the third stage or the adult stage.

One important biological phenomenon that occurs on cadavers are a succession of organisms that thrive on the different parts. E.g. beetles that specialize on bone, will have to wait until bone is exposed. Predatory rove beetles or parasites that feed on maggots will have to wait until the blow flies arrive and lay their eggs.

The succession on cadavers happens in a fairly predictable sequence and can be used in estimating time of death if the body has been lying around for some time.

There are several things to note about this table:
The first groups to arrive is blow flies, followed shortly by staphylinids. As putrefaction develops, more groups arrive at the scene, with most groups present just before the body is drying out due to seepage of liquids. After the body is drying out, dermestids, tineids and certain mites will be the dominant animal groups on the body, and blow flies will gradually vanish. Note also how the fauna changes in the soil. This can also be used to estimate time since death.

Succession data can be incorporated in a database, and when the forensic entomologist investigates a case, he can use the taxa found on the body as input, and get an estimate of the time of death as output.


Day number
Taxa 1 2 3 4 5 6 7 8 9 10
A 1 1 1 0 0 0 0 0 0 0
B 0 0 1 1 1 1 0 0 0 0
C 0 0 0 0 1 1 1 1 1 1
D 0 0 0 0 0 0 1 1 1 0
E 0 0 0 0 0 0 0 0 1 1

The hypothetical table above shows the presence (1) and absence(0) of five different taxa (A, B, C, D and E) over ten days. The tabulated data is usually obtained from decay studies done on pigs or other animals.
Let’s say the investigator finds taxa C, D and E on the remains. From the table above, we can see that taxa C occurs on the cadaver from day 5 to day 10, and taxa D from day 7 to day 9, and taxa E from day 9 to day 10. By studying the overlap, the entomologist estimate the PMI to be about 9 days.

Several insects are specialized in living in very decayed dead bodies. One example is the cheese skipper, Piophila casei, where the larvae usually occurs 3-6 months after death. The cheese skipper is a well known pest of cheese and bacon worldwide, and has a cosmopolitan distribution. Adult cheese skippers may occur early after death, but larvae occurs later. The earliest observation on human remains are when the body is two months, and this was under excellent summer conditions. In 1898, Potter examined 150 graves, and found remains of P. casei in ten of them. These graves were from three to ten years old and three to six feet deep.

In temperate regions dead bodies often appear in spring, after the snow is gone. The forensic entomologist and the forensic pathologist must then try to determine whether the death occurred during the winter or before the snow set in. If the death occurred before November, it is possible to find dead insects in and on the body. By analyzing the dead insect fauna, and estimating when the insects probably died (this can be found by looking at meteorological records). Another hint is when the different adults stop flying before the winter. For example: we have had some cases where the bodies have been found in the spring. In one case we found dead third stage blow fly larvae in the back of the mouth. The blow fly larva was of an species that is flying from May to October. It was from this concluded that the eggs probably was laid during October, and since it was relatively few larvae, probably late in October. In another case, we found several live insects on a dead body, and also many dead third stage larvae. The dead larvae was found on the stomach, the arms, the shoulders, and inside the head. We concluded that the live insects had colonized the dead body in the spring, and that the dead larvae had died during the winter. Based on the widespread occurrence of the larvae, we had to say it was likely that the body was colonized before October, probably in September.

If the death occurred in the winter things become difficult in outdoor settings, as very few insects are active in the winter. It is reported that larvae of the winter gnat, Trichocera sp. can develop on carrion in the winter. By estimating the age of these larvae, if present, it could be possible to estimate the PMI.


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