Forensic Entomology ( Chye Shu Yi, Jefferson Tan)

Forensic entomology = science and study of insects and other arthropods with law-related applications by accredited person No need for forensic entomologists in Spore because body decomposes within 3-5 days due to weather/ environ + size of Singapore + efficient police force; impt for temperate cntries Applications: Urban Forensic Entomology o E.g., pest infestations, civil litigation Stored-product Forensic Entomology o Infestation of commercially available foodstuff Myiasis o Insect larvae feeding on necrotic/living tissue o Useful to prove caregiver negligence/ill-treatment Maggot Debridement Therapy o Maggots applied for 2-3 days to non-healing wounds when modern methods have failed o Maggots dissolve and eat dead and infected tissue, kill bacteria and speed up healing rate o Used in Tan Tock Seng and approved in Msia o Discovered for use during WW2 Taxidermy/ bone cleaning o Dermestes beetles feed on protein-based products, leaving behind clean bones o Beetles are potentially pests, e.g. in museums Medico-legal/ medico-criminal entomology* o Anthropod involvement in criminal/ legally precarious situations, e.g., rape, abuse, trafficking (cond) Through yield of DNA in victims blood To determine ante/post-mortem events, e.g., time since death Toxicology analysis of human tissue after having a maggot blend

History of Forensic Entomology Song Ci (1235): Insects attracted to sickle with invisible tissue remnants Francesco Redi (1668): Refuted spontaneous generation of life hypo Bergeret (1855): First westerner to use insects as forensic indicator Megnin (1883, 1898): Published Faunes des Tombeaux & Cadavres Reinhard (1881): First German forensic entomologist Aldrich (1916): Monograph on Sarcophagidae family Knipling (1935, 39): Descripn and keys to first instar of flesh fly maggots Hall (1948): Monograph Blowflies in North America: identify adult and mature larvae of most species in the family Hall, Townsend, Goddard, Lago: Distributional info on forensic flies Meek et al. (1983): Initial guide to forensic pathology Greenberg (1985): rearing data of forensically impt species to deg hr Vincent et al. (1985): Biblio of all pubs dealing with medico-crim ento Smith (1986): First textbook: A Manual on Forensic Entomology Liu, Greenberg (1986): SEM for acc ident of crit larval and pupual stgs (1990): Procedural guide Entomology and Death published for crime scene investigators and other forensic specialists

Life cycle of insects Mates upon emergence from pupa Feeds on protein from body fluids Moves from crpse to suitable pupation site No feeding Trnsforms to pupa Clumps of 300 4 days to pupa Lives in puparium 10 days to adult Feeds on fluids exuded from body Migrates into body 1 day to first moult Move in maggot mass 1 day to 2nd moult 1 day to hatch Lays eggs on corpse 2 days to egg laying

Two KEY Concepts: Temperature at the micro-habitat level affects development time o Optimal temp = best rate of enzymatic action Ecological succession = diff group/species over time due to changes by each o Pioneer secondary etc.

Factors affecting development & succession: Geography (Temperate v Tropics) Seasons (Summer v Winter) Exposure to sun (Sun v Shade) Urban (buildings) v Rural Buried bodies (depth of burial) Bodies in water/ veh/ enclosed spaces Hanged/ burnt/ wrapped bodies Drugs, toxins, food eaten before death

Moves in mass Greatly inc in size 2 days to prepupa

NB: Many drugs affect insect development: Malathion(suicide)=delay colon in mouth Cocaine=accel dev of some Sarcophagids

Factors affecting insect infestation: Physical: water, depth buried, wounds Chemical: embalming, insecticides, lime Climatic: heat, cold, wind, rainfall Animals: birds, mice, rodents etc.

Corpse

Usage by Consumed by

Characteristics of different families Incidentals (e.g., springtails, spiders, mites) Use corpse as an extension point Omnivores (e.g., ants, wasps and some beetles) Consumd Consume corpse by and decomposers by Muscidae (houseflies) Adults have spongy mouthparts; feeds on fluids/ solid food liquefied by their fly saliva Larvae usually dung feeders Calliphoridae (blowflies) Attractive blue/green metallic colour Relatively large in size Picks up faint traces of (decay) odour from up to 20 km from birth-place Larvae feed using mouth hooks

Sarcophagidae (flesh flies) Stripey-backed or chequered, with bright red eyes Histeridae (Hister beetles) Shiny black or metallic--green beetles with introverted heads Staphylinidae (Rove beetles) Elongated beetles with small elytra (wing covers), large jaws Silphidae (carrion beetles) Large in size Nitidulidae (sap beetles) Small, ovoid, knobbed antennae

Broken down by Decomposers (e.g., vertebrates, invertebrates, fungi, protists (bacteria)) Imptly, necrophagic insects Attacked by Predators/parasites (e.g., beetles, true flies, wasps) Attacks immature flies

Musca domestica feeds on exuded fluid from corpse

Blowflies (Calliphoridae)

Flesh flies (Sarcophagidae)

Hister beetles (Histeridae) Adults lay eggs in corpse; larvae feed on blowfly pupae

Rove beetles (Staphylinidae) Adults are early visitors to a corpse and eat all larvae, incl. other predatory fly larvae

Carrion beetles (Silphidae) Usually hide under carcass Cannot survive alone; will die w/o access to maggots

Sap beetles (Nitidulidae) Feed mainly on decaying veg matter, overripe fruit, sap, oozing fluids from decaying body, carrion

Pioneer flies (muscidae + calliphoridae) arrive Eggs/ larvae laid on body Larvae + adults feed on fluids exuded from body Later enter body through nat openings/ wounds but finally feed ovr bod as tissues decay

Sarcophagidae arrive, competes with blowflies for best laying sites; can be seen kicking other flies away.

Generally hide under corpse during daylight and They lay eggs in corpse and their larvae are at night, enter maggotinfested part of corpse to predatory as well capture & devour maggots

Cheese flies (Piophilidae)

True flies (see below)

Coffin flies (phoridae)

Hide beetles (Dermestidae) WHEN CORPSE IS DRY Only beetles with enzymes req. to break down keratin (part of hair and nails) Feed on dried skin, tendons and bone left behind

Ham beetles (Cleridae) Larvae & adults predatory Common in later stages of decomposition of carcass Larvae feed on dried fat & pupate in empty pupal cases of other flies after sealing e opening with silk

Piophilidae BUTRYIC FERMENTATN attracted to corpse during later parts (particularly butyric fermentation) Can dig into ground! :) Characteristics (contd)

Australophyra rostrata attracted to corpse at later part of decomposition after maggots of blowflies have disappeared but before corpse is fully dry. Maggots feed on smooth maggots

Phoridae appears when butyric fermentation begun and when corpse is starting to dry Commonly found in buried human coffins; CAN DIG!

Piophilidae (cheese flies) Attracted to cheesy smells Larvae can flex and skip into the air Phoridae (coffin flies) Minute flies Able to dig through cracks in the soil! Calliphoridae (screw worm) Adult has bright green colouration Parasite in warm-blooded animals Scarbaeidae (scarab beetles) Beautifully coloured, large and hard, highly polished

Dermestidae (Hide beetle) Hairy larvae Cleridae (Ham beetles) Often have a metallic sheen or are colored red or yellow. Trogidae (carcass beetles) Largesized beetles with very thick exoskeletons and uniform dark colouration.

Screw worm (Calliphoridae)

Scarab beetles (Scarabaeidae) Arrives when body is dry

Carcass beetles (Trogidae) Last beetles to inhabit carcass Feed on dried remains e.g. skin, ligaments Larvae live in vertical burrows under carcass when they are not foraging

Puparium left behind remains for extended period of time and can be used to establish max post-mortem interval estimation

OTHER INSECTS

Mites

Parasitic wasp

Mites

Moths

Gamasid mites e.g. Macrocheles common in early stages of decomposition

Eggs laid inside larvae/ pupae of other flies When the eggs hatch, larvae feed on maggot/ pupa and eventually kill it. Wasp larvae then pupate inside maggot/pupa and emerge as adult wasps

WHEN CORPSE WHEN UNDERGOING CORPSE BUTRYIC IS DRY FERMENTATION

Tyroglyphid mites feed on dry skin in later stages Some mites and carrion beetles have symbiotic lifestyles

Feed on mammalian hair during larval stages Adult moths lay eggs on carcass after all other larvae done with it Thus tineid moths are final animals contributing to decomposition of carcass

NB: Moths and dermastids are substitutes: dermastids more resistant to cold

Collection of insects 1) Adult flies and beetles are fast-moving & leaves scene rapidly once disturbed o Therefore must collect as soon as possible Collected insects placed in mouth of killing jar to immobilize them before transferring to vial of 75% ethanol (preservation) o Beetles - collected with forceps/gloves & placed directly into 75% ethanol 2) Larval specimens Search for presence of eggs Search for and collect the longest larvae Collect representative sample of 50-60 larvae from maggot mass If more than one area of colonisation then each area should be treated separately Collected specimens must be properly labelled Label should be placed in alcohol with specimens o Geographical location o Date and hour of collection o Case number o Location of body where removed o Name of collector Duplicate label should be made and affixed to exterior of vial 3) Live insects Duplicate samples should be made for live specimens Placed in specimen containers along with food substrate / moistness / airholes This container should be placed in a slightly larger container with material in bottom to absorb leaking liquids Entire container put in shipping container to ship out that night 4) Soil samples Soil and litter samples - taken immediately under body & frm surroundings Collect leaf litter and debris down to exposed upper surface of soil o Make separate collection from about first 2-3 inches of topsoil o Soil collection area should be about 4-6 inches square o Taken from under head, torso and extremities 5) Temperature data Ambient air temperature = chest height, therm in shade Maggot mass temperature = therm into larval mass centre Ground surface temperature Interface temperature between body and ground Temperature of soil directly under body Weather data (rain/sun?)

Basic Principles Thermal conditions at scene will fluctuate Data from growth at constant temp- divide periods considered into short intervals Apply model of development closest to mean temperature during each period Adjust time period to fit circumstances of site where body was found Accumulated Degree Hours = time x temp Post Mortem Interval (PMI) Provides a minimum because offspring not deposited on host and many factors influence insect development and succession Estimating PMI Collect and preserve sample prior to estimating PMI Rear others to adult stage for identification Use known developmental times at constant temperature

Making calculations 1. Determine temp history at crime scene to calculate av. temp that body exposed to

2. Rear maggots to adulthood to identify species and time to pupae

3. Estimate time of egg laying Knowledge of development rate at rearing temp = estimate age of maggots when body found Knowledge of development rate at av crime scene temp = count back to determine date of egg laying = latest time at which body has died Degree days Total degrees of heat above lower threshold accumulated in 24 hour period (total accumulated degree days) Degree hours Total degrees of heat above lower threshold accumulated in 60 min.

Problems / Controversies Carrion fly larvae can have a metabolic & feeding rate greater than most immature insects. Large number of larvae in the same location can generate a temperature substantially higher than ambient = maggot mass effect. Larval growth is inhibited as lethal high temperatures are approached. Difference between ambient and maggot mass temperatures is likely to be greatest during cold weather (Not applicable to the Tropics). Also, the oldest larvae on the body may have developed prior to elevation in temperature. Unknown insect fauna of the region.

1. Fresh/ Initial Decay (Days 0-3) Commences at death, ends when bloating is first evident. Intestinal bacteria, protozoans and nematodes; digestive enzymes out of ctrl o Breakdown protein and carbohydrates Blowflies and house flies lay eggs in openin - eye, nostrils, mouth, anus/ genital openin Eggs hatch/ move into bod within 24 hrs Complete life-cycle in 2-3 weeks Longer at low temps 2. Bloated Stage/ Putrefaction (Day 4-10) Gases produced by anaerobic bacteria inflate the abdomen. Seepage of fluids, larval activity speeds up, decomposition speeds up Smells emanating attracts more insects Latter-arriving flies & beetles= predators, feed on maggots + decaying flesh Parasitic wasp lay eggs in maggots + pupae

3. Decay Stage/ Black Putrefaction (Days 10-20) Abdominal wall breaks - gases escape. Carcass deflates. Several gens of maggots present, some fully grown, they migrate from the body and bury themselves in the soil where they become pupae Predatory maggots more abundant at this stage; pioneer flies cease to be attracted to the corpse Predatry beetles lay eggs in crpse,larvae eat decaying flesh Parasitic wasps become more common 4. Post-decay Stage / Butyric Fermentation (Days 10-25) Remaining flesh removed over this period; body dries out, emitting a cheesy smell attracts new organisms Body now more suitable for chewing mouthparts of beetles Beetles feed on skin and ligaments mostly larvae o Adults fed on body during earlier stages Cheese fly consumes remaining most flesh In dry habitats, remains are skin, cartilage, and bones. In wet habitats, wet, viscous material in the soil under the remains. 5. Dry Stage/ Dry Decay (Days 50 365++) Mainly bones and hair remain. Odor is primarily that of normal soil and litter. Can last several months to years. Moths, dermastes, bacteria, micro-organisms etc. feed on hair/ keratinous components Mites feed on bacteria (remains on body as long as traces of hair remain.