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Maggots and Time of Death Estimation Slide 2 Slide 3 Entomology is the Study of Insects Images from: www.afpmb.org/military_entomology/usar myento/files/ArmyEntomology.ppt Slide 4 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 Slide 5 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) Slide 6 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 Slide 7 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. Slide 8 If it werent 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 Slide 9 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. Slide 10 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 corpses 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 Slide 11 Image: http://www.nlm.nih.gov/visibleproofs Slide 12 12 Fly larvae - Maggots First-instar Second-instar Third-instar Slide 13 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 Slide 14 Temperature Stiffness Time of death Warm Not stiff Dead less than three hours Warm Not stiff Dead less than three hours Warm Stiff Dead between 3 to 8 hours Warm Stiff Dead between 3 to 8 hours Cold Stiff Dead between 8 to 36 hours Cold Stiff Dead between 8 to 36 hours Cold Not stiff Dead in more than 36 hours Cold Not stiff Dead in more than 36 hours Slide 15 Body TemperatureStiffness/ Rigor MortisTime of Death Warm Slide 16 Temperature Stiffness Time of Death Slide 17 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 Slide 18 Calculate the heat/thermal energy (accumulated degree hour) required for each stage of the Green Bottle Flys 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. Slide 19 Image: http://www.nlm.nih.gov/visibleproofs Slide 20 FromToTempHoursADH Cumulative ADH Egg1 st Instar70 F2323 x 70= 1610 ADH 1610 1 st Instar2 nd Instar70 F2727 x 70= 1890 ADH 1610+ 1890 2 nd Instar3 rd Instar70 F2222 x 70= 1540 ADH 1610+1890+ 1540 3 rd InstarPupa70 F130130 x 70= 9100 ADH 1610+1890+ 1540+9100 PupaAdult Fly70 F143143 x 70= 10010 ADH 1610+1890+ 1540+9100 +10010 Slide 21 Fresh Bloat Decay Post-decay Dry (skeletal) Slide 22 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. Slide 23 Swells due to gases produced by bacteria Temperature rise of the corpse Flies still present Slide 24 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 its original mass. Slide 25 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. Slide 26 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. Slide 27 This 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. Slide 28 Both 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. Slide 29 The pigs were kept at -80C 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. Slide 30 Pigs with a genetic line of a minimum of fifty percent Yorkshire. They were 8-10 weeks old and weighed approximately 40-50 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. Slide 31 Daily 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. Slide 32 Using 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. Slide 33 The 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. Slide 34 Spiracles are incomplete Third-instar larvae Slide 35 Spiracles are complete Third-instar larvae Slide 36 Flies began to arrive within minutes of pig placement however, laying of eggs was delayed 12-18 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. Slide 37 At 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. Slide 38 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. Slide 39 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. Slide 40 The graph shows an elevation for maggot mass temperatures over ambient The fluctuation in ambient temperature induced elevated maggot activity which is consistent with other similar experiments. Sunlit Pig Slide 41 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 Slide 42 Shows a gradual increase then decrease for the Phormia regina The maggots feed and grow to a certain point when they begin to leave the carcass to find a safe place to pupate. Slide 43 Two peaks for the Phaenicia Infers two generations for Pig 1. Slide 44 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. Slide 45 Two 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. Slide 46 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. Slide 47 ADH for Pig 1 was calculated as 4885.2 after nine days. ADH for Pig 2 was calculated as 4488.6 after nine days. These can be used to determine PMI for carcasses found in this area in similar conditions. Slide 48 Slide 49 The End