Module 3.

Soft Tissue Decomposition and Despite autolysis being the first process
DNA to start after death, both autolysis and
Decomposition Research putrefaction occur simultaneously.
 Vertebrates (has backbone) have Furthermore, different rates of autolysis
significant ecological effects when are observed varying metabolic activities
they decompose. and lysosome counts across cells.
Though a small portion of biomass, there
is a significant effect on the For instance, cells between the
environment. Their decomposition results gastrointestinal degrade first due to
in a nutrient-rich hot spot that supports their metabolic activity and lysosome
a diverse ecological community including count.
scavengers and microbes called necrobio.
Understanding the carrion (dead and Stages of Soft Tissue Decomposition
decaying flesh) and human bodies
decompose is important for archaeology Fresh
and forensic anthropology. It provides  Decomposition has begun.
information to questions about PMI,  Most changes are not visible except
location of burial, cause of death and potential clouding of the cornea.
more.  Minimal maggot activity but eggs
from flies maybe visible.
 Animal proxies (such as pigs) have  Other changes like pallor
their advantages in controlled lab (w3hitening of the skin), muscle
experiments but may not perfectly relaxation, and drying of the
represent human decomposition. corneal epithelium are common.
 Human-focused research (such as  Mortises (rigor, livor, algor) can
human body farms) facilities can begin in this stage.
provide more accurate insights, but
they face ethical, legal, and Early Decomposition (Active Decay)
logistical hurdles.  It begins potentially 24 hours after
death
Dr. William Bass  Includes autolysis and putrefaction
 Key figure in Forensic Anthropology characterized by skin slippage,
and Forensic Taphonomy fluid-filled blisters, hair loss,
 He established the first Forensic and bloat due to the proliferation
Anthropology Outdoor Decomposition of bacteria.
Research Facility in 1981 due to his  Color change in the body is visible
struggle to identify the PMI of the and becomes noticeable due to
preserved embalmed body enzymes and microbial catabolism
 Fluids from the decomposing body
Autolysis and Putrefaction create a nutrient-rich area
 Autolysis is the process where cells surrounding the body called cadaver
break down themselves after death decomposition island.
due to enzymatic self digestion. It
starts within minutes after death Advanced Decomposition
when cardiac arrest inhibits oxygen  This stage is characterized by
and blood circulation throughout the extensive mass loss due to continued
body. This would lead to a declined insect activity
intracellular PH. Anaerobic  The weight/mass loss of the cadaver
conditions disrupts ATP production is caused by the maggots eating the
and biosynthesis leading to a soft tissue.
further drop in cytoplasmic PH. This  The body takes on a deflated
condition compromises a cellular and wrinkled appearance with initial
organellar membrane integrity skeletal exposure usually in the
causing the release of hydrolytic head.
enzymes from cell lysosomes. These  Presence of arthropods (diptera).
enzymes degrade cellular structures Flies were replaced by beetles
and macromolecules eventually
 Soil surrounding the cadaver is rich
spilling cellular contents into the
in nutrients like carbon, nitrogen,
intercellular spaces.
magnesium, calcium, phosphorus, and
potassium
 Putrefaction involves the
decomposition of the body internally Skeletonization
due to the unchecked growth of  Characterized by more than 50%
symbiotic got bacteria in an skeletal exposure of decomposing
anaerobic environment after death. remains.
Large macromolecules like lipids,  The environment surrounding the body
proteins, nucleic acids, and reverts to its original state
carbohydrates are catabolized by  Bone continues to interact with the
bacterial fermentation leading to the environment through process known as
production of organic acids and gases. diagenesis leading to chemical
These gases causes bloating in the and/or microbial bone degradation.
abdomen and other body regions. Alcohols
like ethanol, methanol, prophanol, and Adipocere and Mummification
buthanol were also byproducts of this  Adipocere formation and
fermentation. mummification, can occur at any
point during soft tissue
 decomposition and they can 1. Presence of Clothing (prolonged
complicate estimating the PMI moisture)
 Adipocere- aka grave wax, results 2. Trauma (might influence decomposition
from the hydrogination of free fatty patterns)
acids to saturated fats 3. Acidity or PH
 Mummification- typically occurs in 4. Burial Practices
dry environments under extreme 5. Scavenging
temperature holding decomposition 6. Seasonality
when moisture loss is complete 7. Geography
8. Deposition Environment
Extrinsic Factors
External Factors that has influence Burials
towards the decomposition Tend to slow down decomposition by
moderating temperature changes and
 Temperature- most important factor limiting to scavengers and insects
regulating decomposition rates, it The conditions of burial such as depth,
affects chemical and biological soil, texture, PH among others always
activities including enzymatic influence the rate and trajectory of
activity, reaction efficiency, decomposition.
insect activity, and microbial
activity. Insects will have a hard Decomposition Location
time have access to the cadaver if Where the remains decompose significantly
the temperature is 13 degrees influence the decomposition. For
celsius or lower. The colder the instance, indoor decomposition usually
area, the lesser the flies. Warmer results in smaller temperature
areas increases the insect activity. fluctuations and reduce access to insects
and scavengers which leads to delayed
General Rule: The rate of enzymatic insect colonization and prolonged periods
activity increases by two (2) or of desiccation.
three (3) times for every 10 degrees
celsius increase in temperature. Intrinsic Factors

Body Size
 Thermal Energy Units
 Aka Cadaver Mass
Researchers use accumulated degree
 Can influence decomposition and how
days (ADD), accumulated degree hours
its effect has been studied in
(ADH), cumulative degree hours (CDH)
various ways.
to account for and normalize the
impact of temperature on  Increased mass leads to larger
decomposition. These thermal energy concentration of various chemicals
units are often incorporated in that can impact PMI
forensic entomology.  Smaller carcasses, faster rate of
decomposition.
Insect Activity  Studies have shown no significant
The second most important rate-limiting differences in decomposition of
factor. It influences the decomposition human subjects of different mass or
process. weight. While, others have reported
Diptera (flies) - Coleoptera (beetles) more rapid decay in larger cadavers.
they are primarily responsible for soft
tissue decomposition. The presence of Sex
insects can drastically accelerate No effect on the rate of pattern of soft
decomposition rates. tissue decomposition

Moisture Age
The availability of water through Harder to discern due to skewed
humidity, precipitation, soil moisture, demographic in human decomposition
tissue moisture affects both insects and research facilities.
microbial activity. There are certain
blowflies species that are attracted to
oviposition sites that are in contact Human Microbiome
with water. Moisture also affects Potential Influence of Human Microbiome
microbial activity by mediating the on individual variability of
diffusion of oxygen, nutrients, and decomposition:
wastes within the environment.
There are differences between
Microbial Activity individuals, the succession of microbes
Research in recent years has shown that seems to be consistent and producible.
microbes play a significant role in
decomposition. Drug-Use can also affect the
Enteric bacteria are crucial for decomposition. Drugs can kill microbes.
putrefactive processes in early
decomposition and soil microbes also
contributes significantly to PMI
decomposition.
Initial PMI Estimation
Additional Factors Methods on Initial PMI Estimation are
applied within the first 24-48 hours
after death and usually more accurate Volatile Organic Compounds
than longer PMIs. Decomposition process produces VOCs which
Methods emit a characteristic odor.These VOCs can
1. Mortises (Rigor, Algor, Livor) and be profiled and potentially used to PMI
they examine super-vital reactions. estimation.

Insect Colonization (ADD, ADH, CDH) These methods offer potential for
estimating PMI more accurately and
However, this method have limitations objectively than the anthropological
because the conditions which the methods. Further research must still be
observations were made do not always validated before they can be carried in
correlate well with the actual conditions Forensic Investigations.
at the death scene.
Soft Tissue Change DNA in Skeletal Remains
Throughout decomposition tends to be more Crucial in both anthropological and
qualitative than quantitative, since it forensic contexts
is qualitative, the method has challenges The analysis of DNA from skeletal remains
including inconsistencies across are crucial because DNA from bones and
different environments. teeth offer insights into fast
individuals and populations. (Under
Scoring System Archaeology)

Total Body Score

It has been developed to quantify soft When soft tissues are unrecognizable, the
tissue decomposition across three regions examination of skeletal remains is often
(head & neck, torso, and appendages). the only way to extract valuable
This method has shown high inter-observer information.
reliability, but has also been critiqued
by its statistical approach and other When it comes to DNA Analysis, there are
flaws. Most reliable even if flawed. challenges such as:
Decomposition has also been measured via 1. Condition of DNA on skeletal remains.
an assessment of the percentage of mass It can be highly variable, with low
over time (m/t) amounts of DNA changes due to
Best fit sample: Universal Postmortem decomposition posing challenges for
Interval subsequent genetic analysis.
2. Humanade Identification and DNA -
Estimation Model- developed by Arpad Vass genetic analysis is n essential tool for
(controversial anthropologist) positively identifying human skeletal
Which measures the percentage of soft remains, involves comparing DNA profiles
tissue decomposition, however, research from unknown remains to unknown profiles
validating this formula shown mixed from missing persons or close relatives.
results prompting a call to develop more 3. PCR Process- is likened to a molecular
region-specific despite these challenges copy machine using enzymes to selectively
estimating PMI using gross soft tissue replicate a DNA segment, sensitive enough
changes continues to be an important goal to replicate even poor quality small
in forensic taphonomy due to its amount integrated DNA samples found in
relatively low-cost and simplicity skeletal remains.
compared to biomolecular methods.
DNA
Methods of Estimating PMI Types of Human DNA in Forensics

Microbiome Nuclear DNA (nucDNA)

Studies have shown that the microbial The human gnome comprises nuclear DNA and
communities facilitating decomposition mitochondrial DNA, for forensic
change in a predictable manner enabling investigations the human nuclear gnome is
the creation of microbial stopwatch of mostly used because of its extensive
death. Promising results. variability across the population which
assist in differentiating individuals.
Lipidomics
Lipids are crucial components of cell The DNA within the nucleus of a cell is
membranes, their degradation could unique to each individual except for
potentially be used to estimate PMI. twins.
Advances in Mass Spectrometry have
allowed the study of cellular lipids on a Mitochondrial DNA
large scale. Some early studies have Located outside cell nucleus within
identified lipids that decrease with some mitochondria, exists in hundreds to
advancing decomposition. thousands of copies within each cell,
increasing the livelihood of obtaining
Proteomics analyzable DNA from the degraded samples.
Large scale study of proteins which also Most skeletal remains are empty DNA due
change in a predictable manner after to its uni-parental inheritance and high
death. Some proteins are known to survive mutation rate, empty DNA is often used to
extreme decomposition environments with a in population analysis and can be
slow rate of decay over time making them geographically specific.
potential markers of PMI.
Y-Chromosome DNA
Y-Chromosome is a part of the nuclear to DNA preservation than the absolute age
gnome with an inheritance pattern similar of DNA sample.
to empty DNA.
Useful in addressing questions about DNA degradation does not follow a
population variability and movement. specific trajectory that can predict the
Not too to be used unlike nucDNA. Y- time since death.
Chromosome methods could be beneficial in However, certain conditions like humidity
male specific identifications. can greatly influence the rate of DNA
degradation.
Diagenesis of DNA in Skeletal Remains
Molecular diagenis of DNA in skeletal Depositional Environment
remains are there are changes that DNA retrieved from bones in cooler
accumulate in DNA due to degradation, environment is often better preserved
modification, and interaction with the than DNA from warmer environments.
environment post mortem.
Geochemistry
This process is immediately following Chemical properties of soil, ground water
cell death, and includes strand breakage, and the immediate environment around the
chemical modifications, and microbial bone significantly impact the biological
attack. diagenesis

Autolysis Factors that Affect the Quality and

Cell death due to the rupture of cellular Quantity of Recoverable DNA from Skeletal
membranes. Releasing enzymes that degrade Remains
cellular structures in DNA. One
significant complicating factor in 1. Processing Techniques
retrieving analyzable DNA from older Several Studies have examined the
tissues is the reduced size of template influence of different methods of soft
molecules. tissue removal or maceration on DNA
analysis. Teeth-based methods were
Strand Breakage generally most effective for preserving
Reduces the DNA molecules to an average DNA, while prolonged boiling or the use
under 200 base pairs in length in older of chemical like bleach were detrimental.
soft tissues and skeletal remains. A
large portion of strand breakage is due 2. Embalming
to autolytic process that occurs soon Embalming fluids could degrade or alter
after cell death, the DNA is then DNA and complicate analysis.
digested by the nucleases enzymes capable
of leaving the bonds holding nucleotides. 3. Skeletal Element
Depending on the type of bone, if it has
Microbes Decomposition better preservation and thus may yield
reliable DNA profiles.
Chemical Modifications
Occur over long periods Bones with better preservation may
provide better DNA samples.
Hydrolitic and Oxidative Damage could
lead to DNA degradation. 4. Bone Type
Compact bone generally yields a greater
Factors that Influence DNA Degradation quantity of DNA than spongy bones.

When it comes to preservation of DNA in 5. Teeth

skeletal remains, bone structures can Highlighted as excellent reservoirs for
offer some level of protection against DNA.
DNA degradation.
6. Area of the Gnome
The DNA becomes bound to hydroxyapatite Type of DNA being targeted for analysis
(part of the bone)providing stability and can have significant implications.
shielding against chemical and Empty DNA which is present in higher
environmental factors causing quantity in a cell is more likely to
degradation. yield analyzable segments. The small size
and circular structure of DNA makes it
Molecular Taphonomy less susceptible to enzymatic
Different molecules degrade at different modification than nuclear DNA.
rate, so depending on various factor such
as the local environment. 7. Technological Advances
DNA degradation is connected to Bone Smaller amounts degraded can be analyzed
degradation.When there is a degradation more efficiently and quickly.
of bone, there is degradation in DNA Laboratories are adapting next generation
because DNA binds two hydroxyapatite in sequencing to conduct more complex gnome
bones, as the inorganic bone components analysis including the inclusion of
degrade, DNA is released and loses its additional genetic markers.
protection.

PMI Complexity of Decomposition

Many studies have shown that the
depositional environment is more crucial
 Complex interplay of biological and
environmental factors in
decomposition and DNA preservation
The local environment plays a key role in
determining the degree of decomposition
or DNA preservation.

 Soft tissue decomposes relatively

quickly under most conditions

 DNA in skeletal remains can be

preserved for extended periods under
the right circumstances.

Understanding these process is critical

in many fields including Forensic
Anthropology.

Perfect Solution for the problem of

researchers: Establish Body Farm

Always collecting Tibia, they produce a

lot of DNA copy from it to study it.