4 Insect BiologyInsects 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 organismsThere 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 crimesDetermination 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 deathTraffic accidents with no immediately obvious causePossible 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 includemovement of the corpsemanner and cause of deathassociation of suspects with the death scenedetection 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 insectsdetermines the size and development of the insectscalculates the growth of the insects and passage through stages of the life cycle in laboratorycompares 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
13 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:TemperatureAccess by insectsDepth of burialOther FactorsChemical-- embalming agent, insecticides, lime, etc.Animals disrupting the corpse
14 Time of Death can be broadly estimated up to about 36 hours Temperature Stiffness Time of deathWarm Not stiff Dead lessthan three hoursWarm Stiff Dead between 3to 8 hoursCold Stiff Dead between 8to 36 hoursCold Not stiff Dead in morethan 36 hours
15 Body TemperatureStiffness/ Rigor MortisTime of DeathWarm
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 windowsBody in box or bagCold temperaturesDeeper 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.
20 Calculating PMI from Accumulated Degree Hours (ADH) ToTempHoursADHCumulative ADHEgg1st Instar70° F2323 x 70=1610 ADH16102nd Instar70 ° F2727 x 70=1890 ADH1610+18903rd Instar2222 x 70=1540 ADH1540Pupa130130 x 70=9100 ADHAdult Fly143143 x 70=10010 ADH+1001024100 ADH
21 Five Stages of Decomposition Fueled by Insect Activity. FreshBloatDecayPost-decayDry (skeletal)
22 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.
23 Bloat Swells due to gases produced by bacteria Temperature rise of the corpseFlies still present
24 Decay Gases subside, decomposition fluids seep from body. Bacteria and maggots break through the skin.Large maggot masses and extreme amounts of fluid.Unpleasant odorLarvae 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.
25 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.
26 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.
27 MethodsThis project took place at the Huntington landfill beginning on September 5, 2003.Two different areas were chosen to deposit two pigs.Pig 1 was laid in a sunlit area.Pig 2 was laid in a shaded woodland area about 100 feet away at an elevation of approximately 20 feet.
28 MethodsBoth pigs were placed in cages constructed of wood and one inch chicken wire that were staked to the ground to protect from predatory animals.Prior to starting the project, great care was taken to prevent insect activity from taking place. After they died, the pigs were individually tied in two black garbage bags, placed in feed sacks, and secured.
29 Methods The pigs were kept at -80˚C in the laboratory. They were placed in plastic bins in order to thaw for 48 hours prior to placement at the landfill.Closed environment was maintained until they were deposited at the site.
30 MethodsPigs with a genetic line of a minimum of fifty percent Yorkshire.They were 8-10 weeks old and weighed approximately pounds.Both died on July 11, 2003 approximately 12 hours apart. One died a natural death and the other was culled from the litter.Both of the carcasses were in very similar condition; there were no breaks, tears or cuts in the skin.
31 MethodsDaily observations were made at both sites throughout the day at 7am, 1pm, 7pm, and 1am.Air, ground, and maggot mass temperatures were taken at each visit and observations were recorded.At 7am and 7pm they also collected maggot samples for analysis and photographed the scene.Observations were noted and samples taken for a period of nine days.
32 MethodsUsing insect tweezers, the investigators collected a number of maggots and dropped the samples immediately into boiling water, to kill the bacteria in the maggots and also to straighten their bodies for easier analysis.The maggot samples were taken from different areas of the body in which there were large numbers present.
33 MethodsThe maggots were then placed into a labeled jar and preserved with 70% EtOH.They also collected interesting arthropods for analysis.All of the samples were labeled and stored for later analysis in the laboratory.
34 Phormia reginaSpiracles are incompleteThird-instar larvae
35 Phaenicia speciesSpiracles are completeThird-instar larvae
36 Results: Fresh StageFlies began to arrive within minutes of pig placement however, laying of eggs was delayed hours.There was already some green discoloration on Pig 2 at the beginning of the fresh stage, possibly due to the fact that it was dead about 8 hrs before Pig 1.72 hrs later, the first signs of bloating occurred, ending the Fresh Stage.
37 Results: Bloat stageAt about 72 hours, noticeable bloating began to occur in Pig 1.However, Pig 2 did not show visible signs of bloating until about 92 hours.The gap between the two pigs might have been even greater if they had both died at exactly the same time.
38 Results: Decay Stage Decay stage started around 102 hours. At this point, the maggots had broken the skin and the pigs had begun to deflate.Decompositional fluids began to seep from the carcass.There was a green froth around the pig and also a dark fluid ring around the body of Pig 1.Maggot activity increased tremendously, and maggot mass temperature reached its high during this stage.
39 Results: Post-decay Stage When the experiment was terminated due to the fact that maggot activity had ceased, the pigs had reached the Post-Decay Stage.They were mostly skin, bones, and hair, but there was some tissue remaining.
40 Temperature is a Factor: Pig 1 The graph shows an elevation for maggot mass temperatures over ambientThe fluctuation in ambient temperature induced elevated maggot activity which is consistent with other similar experiments.Sunlit Pig
41 Temperature is a Factor: Pig 2 The ambient temperature for Pig 2 was more constant because it was in a shaded area.The temperatures for Pig 1 fluctuated more than those of Pig 2.Shaded Pig
42 Phormia Average Maggot Length vs. Time Shows a gradual increase then decrease for the Phormia reginaThe maggots feed and grow to a certain point when they begin to leave the carcass to find a safe place to pupate.
43 Phaenicia Average Maggot Length vs. Time Two peaks for the PhaeniciaInfers two generations for Pig 1.
44 Two Different Maggot Generations These are distinguishable by the length and obvious size difference.This is why we believe there are two peaks in our graph data for the Sunlit Pig.The photograph was taken at a time consistent with the influx at 132 hours.
45 DiscussionTwo different species of maggots were collected over the nine day period.These two species were analyzed at their third instar stages; they were able to determine the difference by comparing their spiracles.The third instar was the only stage that they analyzed; species determination was more evident at this stage of development.They also reared a sample of maggots from each pig for later species analysis.
46 Accumulated Degree Hours ADH may be calculated using temperature and hours.This works because there is direct correlation between temperature and maggot development.These calculations were somewhat approximate but relatively accurate.
47 ADH and Pig ResultsADH for Pig 1 was calculated as after nine days.ADH for Pig 2 was calculated as after nine days.These can be used to determine PMI for carcasses found in this area in similar conditions.