Forensic Entomology: A Complete Overview

black and gray fly: Entomology

Depending on the condition of the corpse and the circumstances under which it is found, different approaches are used to calculate the post-mortem interval. Forensic pathologists and scientists from all over the world have been baffled by how to rule out foul play in deaths that are not considered natural. As a result of ongoing forensic science research, new techniques for determining the cause and manner of death are constantly being developed. To ascertain the reason, manner, and more especially the duration of the death, numerous techniques have been developed. When it comes to putrefied corpses discovered in various environments under enigmatic, unidentified circumstances, forensic entomology is one such field that helps in determining the period since the death.

With the introduction of new molecular-level techniques, such as DNA analysis and identification based on entomological data that have changed over time, the field continues to advance. For the benefit of scientists, entomologists, pathologists, and crime scene investigators, standards and guidelines for entomological data collection and processing required for involvement in the crime scene must be updated periodically. Processing entomological data must take into account several variables that affect post-mortem intervals. A thorough examination of an insect’s life cycle, it’s family of related species, and reliable, accepted techniques for collecting, reproducing, and identifying it can shed light on the reason(s), manner(s), time(s), place(s), and circumstances of unknown or unnatural deaths.

Forensic case investigators now have a difficult task on their hands because of the complexity of the victims’ identities and the amount of time since the murder. A small number of circumstantial evidence can be quite helpful in reducing waiting time and excluding potential causes and dates of death. Entomology for forensic purposes is one of the circumstantial factors. In cases of unidentified and unnatural deaths, it is nevertheless of the utmost relevance even though it is not a top priority for crime scene investigation. Mass disasters, both natural and man-made, have increased during the previous ten years. In many situations, forensic entomology can be used to accurately calculate post-mortem intervals.

Forensic entomology is the study of insects that live in the flesh of dead humans and animals intending to assist law enforcement in the investigation of criminal cases. It helps with timing from the moment of death till the remains of the deceased are discovered. The time between the colonization of flesh-eating insects and their final developmental or adult stage is referred to as the “entomological post-mortem interval,” a new phrase that is relevant to forensic entomology research. When the standard post-mortem markers or rate methods have worn off and the remains are putrid or unrecognizably altered, an entomological timeline is used.

The scientific study of an arthropod’s colonization of a dead body is known as forensic entomology. This involves research on the different insect species that are frequently found near cadavers, their life cycles, their ecological presences in specific environments, as well as how the insect assemblage varies as the decomposition process advances. Based on how long a specific species of insect spends in a particular developmental stage and how many generations have been created after the introduction of the insect to a specific food source, insect succession patterns are established.

Because flying insects are drawn to a body shortly after death, insect development together with environmental information like temperature and vapor density can be utilized to determine the amount of time since death. The main focus of this scientific subject is the identification of postmortem intervals to assist in death investigations. Toxicology scenarios to identify the presence of drugs, cases of neglect and abuse, and incidents of dry-shelf food contamination have all used forensic entomology outside of killings. Insect assemblages found on a body can also be used to roughly pinpoint a location because some insects may be specific to a given region.

The three subfields of forensic entomology are urban, stored-product, and medico-legal/medico-criminal entomology.

The concurrent time scale needed to calculate the passing of time is provided by entomology. This concurrent approach of forensic entomological timeline becomes a crucial aspect of death investigations if the time since a death has extended more than 72 hours. With improvements in laboratory methods, species identification and time since death may now be determined from an insect’s DNA.

Mitochondrial DNA analysis using the CO1 gene is now used to identify species of their demographic origin and potential relevance to the crime scene. In 2017, Sharma and Singh noted that if medico-legal institutes and labs support forensic entomologists with appropriate DNA settings, the determination of genetic fingerprinting of specimens of flesh-eating insects may be of higher significance. The DNA makeup varies among different insect species. Because insect gene expressions are distinct, forensic entomologists can more easily identify species and calculate postmortem periods.

Entomology has historically been a lengthy and time-consuming field in forensics, but it can be helpful in cases involving criminal activity that is mysterious and out of the ordinary. Up until the 20th century, studies on insects were restricted to the traditional morphological identification of insects about their predictable life cycles according to their class, order, and species. Legal investigators are now more likely to rely on forensic entomological data than in the past due to scientific developments such as using DNA typing to identify insects and barcoding.

Medico-legal significance of insects

The life cycles of insects are regular and predictable, and they have been well-documented in the past. Each species undergoes metamorphosis in a certain order. A reliable way to identify species and estimate the amount of time since death is to use a fixed and consistent entomological timetable. Scientists have a particular interest in forensic arthropods. Until they are found, gathered, and watched as they mature in a controlled environment, the insect succession of the dead remains follows a natural schedule at the crime scene. This guarantees a correct biological timeframe following colonization.

Though the life cycles are fixed, and predictable, certain environmental factors can shorten or prolong the time of maturation of the insects. The most significant variable is temperature and humidity. Though there are some insignificant factors like shade, rainfall and food sources, and drugs.

The faunal succession

Entomology expertise and skills are required for the assessment of putrefaction in the remains and the correlation of these with insects at various developmental phases. Byrd claims that during the first three months, fresh dead matter first attracts blow flies (Calliphoridae). They produce eggs that are like grains of rice. In the initial few months of putrefaction, the dead stuff emits an odor that draws blows and meat flies (Sarcophagidae). Viviparous flesh flies are present. Intriguingly, if the fat in the dead organic matter turns rancid during the next three to six months, it will continue to draw other types of flies. Additionally, dermestid beetles (Coleoptera). The process of devouring flesh continues for another 4–8 months, and mites can be recovered in 1–12 months. If the dead remains go completely dry in 1-3 years there is still the possibility to discover Dermestid beetles, which are found even after 3 years of death.

Collection of entomological evidence

Evidence should be gathered from above, below, and inside the flesh, once the corpse or remains have been found. Adult live flies should be manually caught with nets if they are hovering above a dead body. Insects are stored in separate containers for dead and alive insects. The containers are appropriately labeled and sealed with information about the location, developmental stage, time of collecting, and names and contact information for the scientist collector. The lab facility rears the live, juvenile insects until they are ready for morphological identification.

In cases of child sexual assault, geriatric neglect, and with a history of bed sores and injuries, particular attention should be paid to the phenomena of myiasis in the peri-anal and vaginal areas that exist before corpse infestation. Even harsh weather conditions and flames won’t harm an insect specimen. Extreme flames can shorten the period of insect invasion within the first few days, but they cannot destroy the circumstances for entomological evidence since they do not prevent insect invasion or the ability to calculate the time since death.

For the evaluation of bug specimens collected there, the location of the incident or crime scene is essential. The location of the discovery, its topography, and its climate all significantly benefit legal investigations. The terrain’s full climatic range slightly facilitates the results of fact-checking. Better fact recognition and crime detection are made possible by the efficient collection and manipulation of this fragile evidence.

Estimation of Post Mortem Interval

The forensic entomological clock has two observable parts.

  • Period of insect isolation
  • Time since insect colonization

Methods to determine PMI

While the time since insect colonization is the stage of development of the insect found at the crime scene, the period of insect isolation is the time of the invasion of flesh-eating insects on the dead matter.

Different insect species have different or distinctive morphological growth cycles. Instars are the phases of a larva’s development during which it grows in length, weight, or shape. After feeding, the larvae seclude themselves in the ground or other dark areas to develop into pupae. The breaking of larvae, also known as eclosion when the adult insect emerges, is the last stage of metamorphosis. The local meteorological center’s weather report should also be examined for a thorough developmental study.

Globally, several techniques are utilized to determine the post-mortem interval using inset growth on dead materials.

Series of stages

Insect succession on carrion is influenced by the biogeographical area where the dead materials are present. The minimum and maximum time intervals since death are determined by these succession patterns. Regardless of how far along the decomposition process is, there are many different species with succession stages that depend on it.

Age-dependent variation of intestinal contents

Insect life cycles can be thought of as exact clocks that begin even minutes after death. The smell of a dead body decomposing draws people in initially. Within a few minutes, the blow flies arrive and lay eggs, which hatch into first-instar larvae. They enter their third instar after molting and enter the roaming stage before pupating. Maggots stop eating as soon as they reach their maximum length, according to a radiological analysis of their feeding habits as larvae. The intestinal filling can be used to estimate the postmortem interval after the anterior intestine is still empty.

On stage invasion

The postmortem interval can be determined from the stage of the insect present on the cadaver by using the following formula:


A = Stage of colonization, B = life cycle stage, C = Correction of climatic factor.

Developmental patterns

Forensic applications can benefit from understanding fly developmental patterns. The postmortem interval is the length of time needed for an insect species to reach the stage of growth. Although they only have brief life cycles and are beneficial in the first three to four weeks after death, blow flies and flesh flies offer a reliable estimate. Post-mortem intervals are estimated as the larvae are raised in a laboratory until they are adult forms. Several elements, primarily temperature and humidity, which affect some factors like the rate of oviposition and maturity, have an impact on insects throughout this time of growth.

Weights of larvae

Larval weight can be determined and used to establish the age of the larvae. Their weights are determined in a laboratory setting with strict controls. A statistical model is developed that links weight distribution to age and estimates the period between egg hatching and the post-feeding period. Inverse prediction or calibration are terms used to describe this approach. If the larva’s age is uncertain, its weight can be compared to the model using inverse prediction to produce a confidence interval. When calculating confidence intervals, the weight of one larva can be considered to have been randomly selected from a population of larvae, with the population being believed to be at the same age and environment.

Since different insect species are found in different regions, distinct models are created for each species. For any particular area, species, or ecosystem, scientists have created their baseline data.

Isomegalen/isomorphen diagrams

At various temperature scales, it is possible to see the entire life cycles of insects as well as their development. This observation may be represented as an isomegalen diagram with temperature on the y-axis and time since egg hatching on the x-axis. The length of the larvae can be shown graphically to estimate an insect’s age. The area between the lines in the graph indicates the morphological phases of insects. The isomorphen diagram shows all the structural stages from egg hatching to the eclosion phase. It can establish the post-feeding larval or pupal age.

Fly eggs

Insects’ propensity for producing eggs can be used to estimate postmortem periods. Insects arrive minutes after a person dies and dine on recently deceased bodies. Data that can be used in genuine forensic cases can be produced by observing the time at which eggs hatch in a laboratory setting with constant temperatures. To establish the postmortem interval, this experimental and developmental data reflects an egg-hatching time within a window of two hours. This can be used to determine a brief post-mortem interval and the emergence of the first instar. These life cycle studies can serve as a foundation for simulation models and are useful for determining their biotic potential.

From insect’s gut contents

After hatching, larvae gorge themselves on the dead carrion and grow quickly in size. Sometimes, because of a lack of food, they stray to adjacent carrion. In such circumstances, DNA characterization of gastrointestinal contents can reveal which species consumed which body. This is crucial proof of the interaction between larval bodies and can serve as a reliable source for calculating the passing of time.

From cuticular hydrocarbons and width

Hydrocarbons are present in the pupal shells; their accumulation and deterioration inside the cuticles are directly related to time. Gas chromatography-mass spectrometry can be used to assess both these metabolic changes and the hydrocarbon ratios that are slowly changing. With these modifications, post-mortem interval estimates using these cutting-edge technologies may become more precise. N-alkanes, methyl-branched alkanes, and dimethyl-branched alkanes are present in the cuticular hydrocarbons of pupal shells. From C21 to C35, the hydrocarbon makeup and length of the carbon chain show notable consistent variations across time.

The abundance of low molecular weight alkanes with even numbers, like n C22, n C24, and n C26, increases steadily and noticeably over time due to weathering. For n C26, for instance, the abundance rises linearly during the course of weathering. With time, the abundance of lower molecular weight hydrocarbons, such as n C26 or less, diminishes significantly.

From accumulated degree days/hours (ADD/ADH) This is another way to calculate the amount of time since a person’s passing. The ADH value is a precise digit of energy hours needed for the development of insect larvae. The degree day or hour idea states that, within the temperature range that is specific to a species, the pace of development is proportional to temperature.

However, this relationship is typically curvilinear at high and low degrees of temperatures and is linear only in between.

Study of wound artifacts; some drawbacks overlooking entomological evidence

Post-mortem insect bite wounds

 Insects like ants and cockroaches appear after the undetectable invasion of fresh dead flesh and begin chewing on the succulent flesh. The abraded tracks left by the gradual chewing on the skin may be mistaken for the obvious vein markings of drug users who deceive investigators or for the trickling of a corrosive down the skin. Some larger punch-out wounds resemble entry or exit bullet wounds. the startling discovery of larvae of flesh-eating insects in the bone marrow, which most likely entered the marrow through vascular foramina in the bone. Dermestid beetles show up and feed on both the corpse and the larvae while the larvae multiply and dry out the flesh. The larder and clown beetles cause bigger pinch-out wounds that resemble firearm wounds on the skin.

Wrongful blood splash marks

Tadpole patterns that resemble blood spatter can be produced when adult flies try to feed on liquid blood tread marks on blood drops splattered around the dead carcass. The direction of the spatter’s tails, the ratio of the tail to the body, and the uneven shape of the blood spots can all be used to rule out this issue.

Misleading investigations

To guide the investigations to a shared conclusion, the participation of all forensic experts is necessary at the crime scene. It’s important to distinguish between antemortem insect invasion and post-mortem insect colonization at the crime scene. Both can frequently be found on the crime scene at the same time, especially when nursing care has been neglected.

In contrast to morphological identifications, which demand an intact specimen, molecular analysis permits identification using fragmentary specimens.

The most prevalent taxa worldwide are insects. We cannot deny the reality that they scavenge the majority of the organic stuff in our environment. It is well-recognized that they play a part in digesting unmanaged dead organic debris. A similar observation may be made in forensics when human corpses are overlooked or dumped indoors, outdoors, or even in remote locales. Usually, strange locales and circumstances fit the condition of the bodies that can be flyblown themselves when such bodies are discovered. In these situations, it is impossible to overlook any evidence at the scene that can trigger more inquiries. In most cases, the bodies are so badly decayed that it is difficult to use standard rate methods to determine when someone died. To interpret the facts, concurrent approaches are therefore considered. Even with the use of these procedures, the techniques and methods for data collection must be appropriate and suitable to get the intended outcomes.


1. Catts EP, Goff ML. Forensic entomology in criminal investigations. Annu Rev Entomol. 1992;37:253–72. [PubMed] [Google Scholar]

2. Amendt J, Krettek R, Zehner R. Forensic entomology. Naturwissenschaften. 2004;91:51–65. [PubMed] [Google Scholar]

3. Goff ML, Lord WD. Entamotoxicology;a new area of forensic investigation. Amer J foren med pathol. 1994;15:51–7.[PubMed] [Google Scholar]

4. Chen WY, Hung TH, Shiao SF. Molecular identification of forensically important blow fly species (Diptera: Calliphoridae) in Taiwan. J Med Entomol. 2004;41:47–57. [PubMed] [Google Scholar]

error: Content is protected !!