1. Maggots and Time of Death Estimation
Forensic Entomology
Maggots and Time of Death Estimation

3. Entomology is the Study of Insects
Images from:

4. Insect Biology
Insects are the most diverse and abundant forms of life on earth.
There are over a million described species- more than 2/3 of all known organisms
There is more total biomass of insects than of humans. of humans.
Insects undergo either incomplete or complete metamorphosis (Egg to larva to pupa to insect)
Larva have a soft tubular body and look like worms. Fly species larvae are “maggots”

5. What is Forensic Entomology?
Forensic Entomology is the use of the insects and other arthropods that feed on decaying remains to aid legal investigations. 
Medicolegal (criminal)
Urban (criminal and civil)
“legal proceedings involving insects and related animals that affect manmade structures and other aspects of the human environment”
Stored product pests (civil) 

6. Medicolegal Forensic Entomology
Often focuses on violent crimes
Determination of the time (postmortem interval or PMI) or site of human death based on identification of arthropods collected from or near corpses.
Cases involving possible sudden death
Traffic accidents with no immediately obvious cause
Possible criminal misuse of insects

7. Postmortem interval (PMI)
Forensic Entomology is used to determine time since death (the time between death and corpse discovery)
This is called postmortem interval or PMI).
Other uses include
movement of the corpse
manner and cause of death
association of suspects with the death scene
detection of toxins, drugs, or even the DNA of the victim through analysis of insect larvae.

8. Forensic Entomology is Applied Biology
If it weren’t for decomposition of all living things, our world would fill up with dead bodies.
When an animal dies, female insects will be attracted to the body. They enter exposed orifices or wounds and lay eggs or larvae.
A forensic entomologist:
identifies the immature insects
determines the size and development of the insects
calculates the growth of the insects and passage through stages of the life cycle in laboratory
compares the growth against weather conditions to estimate time of oviposition

9. Succession of Insects on the Corpse
Estimates of postmortem intervals based on insects present on the remains are based on:
The time required for a given species to reach a particular stage of development.
Comparisons of all insect species present on the remains at the time of examination.
Ecological succession occurs as an unexploited habitat (like a corpse) is invaded by a series of different organisms.
The first invasion is by insect species which will alter the habitat in some form by their activities. These changes make the habitat attractive to a second wave of organisms which, in turn, alter the habitat for use by yet another organisms.

10. Ecology of Decomposition
Necrophages - the first species feeding on corpse tissue. Includes rue flies (Diptera) and beetles (Coleoptera).
Omnivores - species such as ants, wasps, and some beetles that feed on both the corpse and associated maggots. Large populations of ominvores may slow the rate of corpse’s decomposition by reducing populations of necrophagous species.
Parasites and Predators - beetles, true flies and wasps that parasitize immature flies.
Incidentals – pill bugs, spiders, mites, centipedes that use the corpse as an extension of their normal habitat

11. Blow Fly Metamorphosis
Blow flies are attracted to dead bodies and often arrive within minutes of the death of an animal. They have a complete life cycle that consists of egg, larva, pupa, and adult stages.
1st – Adult flies lay eggs on the carcass especially at wound areas or around the openings in the body such as the nose, eyes, ears, anus, etc.
2nd – Eggs hatch into larva (maggots) in hours.
3rd– Larvae continue to grow and molt (shed their exoskeletons) as they pass through the various instar stages.
1st Instar - 5 mm long after 1.8 days
2nd Instar - 10 mm long after 2.5 days
3rd Instar – mm long after 4-5 days
4th – The larvae (17 mm) develop into pupa after burrowing in surrounding soil.
5th – Adult flies emerge from pupa cases after 6-8 days.
Adult
Pupa
Eggs
3rd Instar Larva
1st Instar Larva
2nd Instar Larva
It takes approximately days from egg to adult depending on the temperatures and humidity levels at the location of the body.
Image:
Information: and

12. Examples of Diptera (Flies)
Early Stage Decomposition
Blow & Greenbottle Flies
(Calliphoridae)
Metallic thorax and abdomen
Flesh Fly
(Sarcophagidae) Striped thorax
Life Cycle of a Calliphoridae Fly
Late Stage Decomposition
House Fly
(Muscidae)
Cheese Skipper
(Piophilidae)
Informational Source: Images: Top Left - Middle-Left: Top Right - Bottom -

13. Examples of Coleoptera (Beetles)
Early Stage Decomposition
Early to Late Stage Decomposition
Rove Beetles
(Staphylinidae)
Predator of fly eggs
Clown Beetles
(Histeridae)
Predator of fly eggs
Carrion Beetles (Silphidae) Adults & larvae feed on fly larvae
Late Stage Decomposition
Ham & Checkered Beetles (Cleridae) Predator of flies & beetles;
also feed on dead tissue
Hide Beetles (Scarabidae) Usually the last to arrive
Skin Beetles (Dermestidae) Feed on dried skin & tissues
Informational Source:
Images: &

14. Image: http://www.nlm.nih.gov/visibleproofs

15. Decay Rates Are Variable
Studies of decay rates of 150 human corpses at in the Anthropological Facility in Tennessee (The Body Farm)
Most important environment factors in corpse decay:
Temperature
Access by insects
Depth of burial
Other Factors
Chemical-- embalming agent, insecticides, lime, etc.
Animals disrupting the corpse

16. Time of Death can be broadly estimated up to about 36 hours
Temperature Stiffness Time of death
Warm Not stiff Dead less
than three hours
Warm Stiff Dead between 3
to 8 hours
Cold Stiff Dead between 8
to 36 hours
Cold Not stiff Dead in more
than 36 hours

17. Differentiate between PMI and Time of Death
These may not always equate.
Post mortem interval is restricted to the time that the corpse or body has been exposed to an environment which would allow insect activity to begin.
Closed windows
Body in box or bag
Cold temperatures
Deeper burial

18. Insect species arrive at a corpse in waves like clockwork
Calculate the heat/thermal energy (accumulated degree hour) required for each stage of the Green Bottle Fly’s life cycle.
Possibly the greatest potential source of error in using arthropod successional patterns lies in the collection of speciments.
Must only be done correctly to accurately sample the insects.

19. Image: http://www.nlm.nih.gov/visibleproofs

20. Calculating PMI from Accumulated Degree Hours (ADH)To
Temp
Hours
ADH
Cumulative ADH
Egg
1st Instar
70° F 23
23 x 70=
1610 ADH
1610
2nd Instar
70 ° F 27
27 x 70=
1890 ADH
1610+
1890
3rd Instar 22
22 x 70=
1540 ADH
1540
Pupa
130
130 x 70=
9100 ADH
Adult Fly
143
143 x 70=
10010 ADH
+10010
24100 ADH

21. Initial decay - 0 to 3 days after death
From the moment of death flies are attracted to bodies.
Bacteria and the body's own digestive enzymes (normally in the intestine) spread through the body, contributing to its decomposition.
Without the normal defenses of a living animal, blowflies and house flies are able to lay eggs around wounds and natural body openings (mouth, nose, eyes, anus, genitalia) and any wounds.
These eggs hatch and move into the body, often within 24 hours. The life cycle of a fly from egg to maggot to fly takes from two to three weeks. It can take considerably longer at low temperatures.

22. Flesh flies - Family Sarcophagidae

23. Blowflies - Family Calliphoridae

24. Blowfly Larvae

25. Stage 2: Putrefaction - 4 to 10 days after death
Putrefaction begins; gases produced by anaerobic bacteria; considerable bloating; seepage of fluids;
Larval activity speeds up decomposition - the smells and body fluids that begin to emanate from the body attract more blowflies, flesh flies, beetles and mites.
The later-arriving flies and beetles are predators, feeding on maggots as well as the decaying flesh. They are joined by parasitoid wasps that lay their eggs inside maggots and later, inside pupae.

26. Stage 3: Black putrefaction - 10 to 20 days after death
Abdominal wall is broken and carcass deflates
By this stage, several generations of maggots are present on the body and some have become fully grown.
They migrate from the body and bury themselves in the soil where they become pupae.
Predatory maggots are much more abundant at this stage, and the pioneer flies cease to be attracted to the corpse.
Predatory beetles lay their eggs in the corpse and their larvae then hatch out and feed on the decaying flesh.
Parasitoid wasps are much more common, laying their eggs inside maggots and pupae.

27. Hister Beetles Prey on Blowfly Larvae

28. Stage 4: Butyric fermentation - 20 to 50 days after death
All the remaining flesh is removed over this period and the body dries out. It has a cheesy smell, caused by butyric acid, and this smell attracts a new suite of corpse organisms.
The reduction in soft food makes the body less palatable to the mouth-hooks of maggots, and more suitable for the chewing mouthparts of beetles. Beetles feed on the skin and ligaments. Many of these beetles are larvae. They hatch from eggs, laid by adults, which fed on the body in earlier stages of decay.
The cheese fly consumes any remaining moist flesh at this stage, even though it is uncommon earlier in decay.

29. Stage 5: Dry decay - 50-365 days after death
The body is now dry and decays very slowly. Eventually all the hair disappears leaving the bones only.
Animals which can feed on hair include moths, and micro-organisms like bacteria. Mites, in turn, feed on these micro-organisms.
They remain on the body as long as traces of hair remain, which depends on the amount of hair that covers the particular species. Humans and pigs have relatively little hair and this stage is short for these species.

30. Making Calculations
Step 1. Determine temperature history at crime scene
Extract weather bureau records of maximum and minimum daily temperatures at the weather station nearest to crime scene, over the general period the body has been exposed.
Calculate the average temperature that the body has been exposed to.
Step 2. Rear maggots to adulthood to identify species
Collect a range of maggots, (particularly those that might be the oldest) from the body and rear them (on ox-liver) at constant temperature.
Record time taken until larvae pupate.
Identify fly species from adult characteristics. (For some species, identification from larval features may be possible or they can be identified from DNA samples, if a DNA library is available.)
Step 3. Estimate time of egg laying
Using knowledge of development rate of the particular species at rearing temperature, count back to estimate age of maggots when body found.
Using knowledge of development rate of the particular species at the average crime scene temperature, count back to determine date of egg laying.
This is the latest time at which the body died. (It may have died earlier if there was a delay between death and egg laying. This depends on weather conditions and accessibility of the body to insects).

31. Case Study Establish PMI:
Body found – all larvae collected are Lucilia sericata
The biggest and oldest are mid 3rd in-star larvae
Weather reports from the crime scene indicate the average temp was 21 C°
This species does not colonize bodies for 48 hours after death
Development rate of Lucilia sericata (in hours) at three different temperatures

32. Solution
Use 21 C°
It takes 21 hours to hatch from the egg, plus 31 hours for 1st instar, plus 26 hours for 2nd instar, plus roughly 25 hours for mid-3rd instar
Add 48 hours colonization time
Total: = 151 hours
151 hours/24 hours per day = 6.29…roughly 6 days pmi

33. Fresh Begins at death Flies begin to arrive
Temperature falls to that of the ambient temperature.
Autolysis, the degradation of complex protein and carbohydrate molecules, occurs.
Gasses produced by the metabolic activities of the anaerobic bacteria first cause a slight inflation of the abdomen. The carcass may later assume a fully inflated, balloon-like appearance. Adult and larval blowflies in large numbers attracted to fluids seeping from body, normal soil dwelling fauna depart soil because of seepage of fluids; some muscid flies and ants which can feed on larvae and retard maggot activity.

34. Bloat Swells due to gases produced by bacteria
Temperature rise of the corpse
Flies still present

35. Decay Gases subside, decomposition fluids seep from body.
Bacteria and maggots break through the skin.
Large maggot masses and extreme amounts of fluid.
Unpleasant odor
Larvae beginning to pupate.
Corpse reduced to about 20% of it’s original mass.
Decay Stage - Black Putrefaction (Days 5-11) -- Decay stage begins when the abdominal wall is broken, allowing gasses to escape and carcass deflates. This process is facilitated by feeding activities of larval flies present on the exposed remains. Adult flies start to leave body, mainly larval mass. Carcass begins to assume a blackened, wet appearance, and most of the flesh will be removed by the maggots. Toward end of this period, carcass will begin to dry and beetles feed on drier tissue. Flies start to pupate. Predatory beetles such as rove beetles and histerids come to feed on other insects.

36. Post-Decay Carcass reduced to hair, skin, and bones.
Fly population reduced and replaced by other arthropods.
Hide beetles are dominant in dry environments.
Mite and predatory beetle populations increase.
Postdecay Stage - Butyric fermentation (Days 10-25) -- In dry habitats, remains consisted of dry skin, cartilage and bones. Site for dermestid beetles, histerids, fly pupae, immature and adult rove beetles. In wet habitats, a large quantity of wet, viscous material, termed byproducts of decomposition, was found in the soil under the remains. Site for immature and adult moth flies, sphaerocerid and muscid flies, rove beetles.

37. Dry (Skeletal)
Does not always occur especially if corpse is in a wet region. Maggots will stay longer and hide beetles will not appear.
In wet environments the hide beetles are replaced with nabid and reduviid insects.
The corpse is reduced to at least ten percent of the original mass.
In the last stage (Skeletal Stage), only bone and hair remain.
Dry Stage (Days 25 +) -- This stage is reached when mainly bones and hair remain. Odor is primarily that of normal soil and litter. Some dermestid beetles, histerids, fly pupae, immature and adult rove beetles, normal soil fauna (mites) start to return. Can last several months to even years.

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40. The End