6 Forensic Examination of Animal Hair

Claire Gwinnett*
Department of Forensic and Crime Science, Staffordshire University,
Stoke-on-Trent, Staffordshire, UK

6.1 Introduction 63
6.2 Hair as Evidence 63
6.3 The Use of Animal Hair in Criminal Casework 64
6.4 Recovery, Documentation, Packaging and Storage Methods
for Animal Hair Evidence 64
6.4.1 Recovery of questioned aka target animal hairs 65
6.4.2 Recovery of control aka known hair samples 67
6.4.3 Packaging and storage 67
6.4.4 Documentation of evidence 67
6.5 General Structure of Hair 67
6.5.1 Types of hair 68
6.6 Forensic Animal Hair Analysis 68
6.6.1 Stages of hair analysis 68
6.6.2 Microscopy preparation of animal hairs 70
6.6.2.1 Creating a whole mount 70
6.6.2.2 Scale casts and impressions of the
animal hair surface 70
6.6.2.3 Medulla slides 70
6.6.3 Microscopical analysis of animal hairs 71
6.7 Species Identification from Animal Hair 74
6.7.1 Scale morphology 74
6.7.2 Medulla types 76
6.7.3 Medullary fraction (MF) aka medullary index (MI) 77
6.7.4 Colour banding 77
6.7.5 Root shape 77
6.7.6 Species identification aids 77
6.8 Interpretation of Animal Hair in Casework 77
6.8.1 Conclusions from comparing control and target hairs 77
6.8.2 Transfer and persistence for interpreting animal crimes 78

*Corresponding author: [email protected]

62 © CAB International 2016. Practical Veterinary Forensics (ed. D. Bailey)

 Forensic Examination of Animal Hair 63

6.1 Introduction 6.2 Hair as Evidence

The use of hair as evidence in criminal Hair can be defined as ‘any of the fine
casework is well established, with hair thread-like strands growing from the skin of
being a common type of trace evidence re- humans, mammals, and some other animals’
trieved from crime scenes (Petraco, 1987; (Oxford Dictionaries, 2014). Composed of
Robertson, 1999). Hairs are readily lost the protein keratin, hairs are generally
from individuals, whether human or ani- stable in nature, hence forensic scientists
mal, and these hairs may be transferred are able to use this type of evidence more
during a crime, helping to link suspects to readily in environments which have de-
scenes, suspects to victims, objects to graded other biological matter (Taupin,
scenes or animals to individuals (to name 2004). Hair evidence has been used in all
but a few possible permutations). Edmond types of criminal cases, including, but not
Locard’s Exchange Principle (Locard, 1930), limited to: murder, sexual assault, burglar-
‘every contact leaves a trace’, explains the ies, abuse cases, arson and terrorist inci-
mechanisms for trace evidence transfer at dents (Robertson, 1999). This breadth of use
crime scenes. It is a fundamental principle is partially due to the ease of transfer of
that whenever objects, people or animals hairs between individuals, individuals and
come in contact with each other, an ex- crime scenes and individuals and objects.
change in material will occur. This in- Hairs transferred to highly important ob-
cludes hairs, fibres, glass, paint and any jects from a crime, e.g. a balaclava identified
other particulate evidence. In the past, ani- as being used in an armed robbery, enables
mal hair was disregarded as forensic evidence the wearer to be identified and linked to
and not frequently analysed in forensic la- the scene.
boratories, but now animal hair is common- Hairs used in the analysis of a criminal
place in forensic hair analysis (Petraco, investigation are defined as either ques-
1987), partly due to development in identi- tioned (aka target) or control samples. Tar-
fication schemes and research outlined in get hairs are the extraneous hairs that have
this chapter. been transferred during contact and are the
This chapter aims to provide to veterin- evidential hairs that will provide informa-
arians, personnel who work in animal wel- tion about the case.
fare (e.g. RSPCA officers), forensic scientists, Hair evidence can provide a large range
police officers and anybody who may be of different information beyond associations
tasked to analyse animal hair for criminal between suspects, victims, places and ob-
casework, an overview of forensic hair ana- jects depending on the type and quantity of
lysis. The focus of this chapter is to provide hair available. Hair evidence can provide
an insight into how animal hair evidence intelligence information in the form of sus-
should be reliably recovered, documented, pect descriptions, i.e. hair colour and type,
analysed and interpreted for criminal cases. as well as information regarding how the
Human hair analysis will not be de- person treats their hair (for example, the
tailed in this chapter, but interpretation use of styling products and dyes) and any
methods used when analysing human hair diseases present in the hair. Examination of
that are also applicable for animal hair will the hair can identify whether it is human or
be introduced. Similarly, textile fibres evi- animal and the body area from which it has
dence will not be focused upon, but as ani- been shed. In the case of human hair, the
mal hair may be defined as a type of ‘fibre’, ethnic origin of the individual the hair has
and are used abundantly in textiles such as been shed from may also be determined.
clothing (Wildman, 1961), many of the If nuclear material is present on the root,
underlying principles of recovery, docu- then the potential for DNA profiling can
mentation and interpretation are compar- allow for a more conclusive identification of
able and will therefore be described where the individual, beyond just microscopical
appropriate. analysis.
64 C. Gwinnett

Hair evidence is a transient evidence has been transferred from the dog owner’s
and due to this the evidence will transfer and clothes to a victim of a crime during an as-
persist depending on certain factors (discussed sault. The information that can be obtained
in Section 6.8.2 below). By understanding from an animal hair is the same as in human
transfer and persistence, timeframes of when hair, although some information is not ap-
certain contact occurred can help to recon- plicable, such as ethnic origin. Animal hair
struct a crime scene and understand whether analysis in criminal cases primarily focuses
the evidence has been transferred at the time upon the species identification of the animal
of the crime, thus making it evidentially use- it originated from, although identification of
ful. Hair evidence can also provide informa- a particular individual has been carried out.
tion about any drug use of an individual. In terms of animal-related crimes, including
Any damage to the hair can also provide the wildlife crimes, hair analysis can be widely
investigator with information about the used: for example, illegal fighting involving
crime and potential suspect(s); this includes dogs, cockerels or hogs (USA); intentional
heat, decomposition and fungal damage. poisonings; poaching; carting of deer; ritualistic
Hair evidence, like other evidence, has crimes; hit and runs; cruelty cases; badger bait-
limitations. The lack of acknowledgement ing; import/export of endangered animals; and
of these limitations in the past has led to animal products and bush meat. In rare cases,
miscarriages of justice and led to investiga- animal hair may be utilized as the primary
tors overstating the value of the evidence source of evidence, but more commonly it is
(Taupin, 2004). The main limitations include used as corroborative evidence, analysed ini-
the subjectivity involved in the microscop- tially to obtain intelligence information or to
ical analysis of hairs and the intra-variation justify further, more costly analysis (as a
in hair characteristics seen in individuals; screening tool). Following are examples of
this will be discussed later in the chapter. the types of cases that utilize animal hairs as
Knowledge of these limitations does not evidence (Case Studies 6.1, 6.2 and 6.3).
undermine hair evidence, but enables inves-
tigators to correctly interpret it.
6.4 Recovery, Documentation,
Packaging and Storage Methods
6.3 The Use of Animal Hair for Animal Hair Evidence
in Criminal Casework
It is important for anyone attempting to locate
Hair evidence from animals has been used and retrieve animal hair evidence to be aware
successfully in the solving of many human of the three Rs of evidence; Recognition, Re-
crimes; for example, when domestic dog hair cording and Recovery (Robertson and Roux,

Case Study 6.1 .

A farmer had a prolonged problem with sheep regularly walked in the area and whose dog
worrying, culminating in one of her livestock had been seen chasing the sheep previously
being killed and mutilated. Veterinary exam- was questioned regarding the case, but re-
inations indicated that the wounds were probably fused to allow control samples to be taken
caused by a dog, but this was not conclusive from his dog for comparison. The information
because of potential interference from scav- gathered in the analysis of the hairs was pre-
engers. Hairs found in the wound of the sented in court in order to obtain sufficient
sheep were retrieved and analysed and identi- control samples and allowed a conclusion
fied as being of canine origin, leading to fur- that the hairs from the wound and the dog
ther investigation, including DNA analysis, of could be associated, linking the dog to the
the hairs. The owner of a local dog that was sheep carcass.
 Forensic Examination of Animal Hair 65

Case Study 6.2.

Meat from a London market had been seized Before testing of the meat, police wished to as-
by the police as it was suspected to be illegally certain whether it was from an endangered ani-
imported bush meat (described as gorilla mal or legal livestock such as bovine. Hairs
meat). The meat appeared to have been treated taken from the skin were identified as being
(smoked and charred) but hair was still present from sheep origin, thus eliminating the need
on the skin (this is common with bush meat). for further analysis.

Case Study 6.3.

It was suspected that a racehorse was wrongly clean locations on the racehorse, where there is
administered a drug over a lengthy time period; low variability in growth rate and hairs had
however, because the dose terminated one grown to sufficient length to span the suspected
month earlier, removing the opportunity for timeframe of the dosing. Segmental hair ana-
blood and urine tests. Hair can provide infor- lysis was carried out. This is where the hair is cut
mation regarding type of drugs administered into segments and analysed using a suitable
and drug use history, and, although hair is not as technique, such as gas chromatography, to
accurate in determining concentrations of drugs allow the time of administration to be identified
at a particular point in time (e.g. at the time of a by locating the position of the drug along the
crime or just before death), it can provide a pro- hair shaft. The results indicated the presence of
file of drug use over a long time period. Conse- clenbuterol hydrochloride in mane hairs ex-
quently, hairs were obtained from different tending back in time by 8 months.

2010). As evidential animal hairs are not ne- 6.4.1 Recovery of questioned
cessarily easily seen when examining exhibits aka target animal hairs
or animals, they may not be recognized with-
out appropriate search techniques. Systematic Veterinarians and other individuals en-
searching of objects and animals for hair evi- countering live animals, carcasses or other
dence should include looking for areas where objects potentially containing animal hair
hair transfer is most likely to have occurred evidence need to know how to retrieve
during the incident rather than through inno- these transient samples reliably and effi-
cent means, e.g. hairs found in a wound of an ciently before they are lost. The acronym
animal suspected of being involved in dog GIFT (Get It First Time) is a principle that
fighting is more evidentially valuable than if anyone attempting to retrieve evidence
found just on the collar of the dog. To identify should abide by and an appropriate method
whether any hairs found are evidentially valu- must be chosen for the recovery of any
able, information such as whether the animal questioned hairs (Robertson and Roux,
or object could have come in contact with any 2010). The retrieval method used must
other animal should be sought, and if hairs allow all target hairs to be recovered, with-
have been taken from a scene, e.g. trailer or out loss or contamination. The recovery
house, knowledge of animal access since the method chosen depends on a number of
crime is required. Without correct recording of factors: surface type, surface area, presence
any animal hair evidence, then the evidential of debris and whether the hairs are loose or
value may not be fully realized and, in the worst embedded in the surface in which they are
case, may not be admissible in court. Finally, found. Table 6.1 outlines the seven methods
without appropriate recovery methods, poten- that are accepted techniques for the re-
tial hair evidence will be left behind. trieval of hairs.
Table 6.1 . Methods for the retrieval of animal hairs.

66
Retrieval
Method Description of Use Preferred Surfaces Advantages Disadvantages

Tape lifting Sticky tape is gently placed on the surface, Any dry porous or non-porous Able to capture multiple Hairs must be dissected from the tape to
removing any surface hairs. Tape is then surface, especially useful for hairs at a time and also allow for further examination due to
placed upon an acetate sheet to preserve smaller surface areas, e.g. inside know from which part of the need to encapsulate hair in a
the evidence and allow for searching of pet carrier. an object the hairs have medium of similar refractive index (RI).
(Choudhry, 1988). been retrieved from. This is time-consuming. A new tape,
called Easylift, has been introduced
that removes the need for dissection
(Jackson and Gwinnett, 2013).
Tweezering Use of clean tweezers to remove obvious All surfaces but usually only when Good for when hairs are Time-consuming for large areas.
hair evidence from surfaces. hairs are found in prominent embedded within a
positions, could otherwise be substance or object, e.g.
easily lost and/or when the exact mud or a wound.
location of the hair is required to
be known.

C. Gwinnett
Vacuuming Use of vacuum filters which attach to a Dry, large surface areas, e.g. pet Quick for retrieving large More time-consuming than other methods
vacuum cleaner and use suction to bedding, large pet carriers. amounts of hair. Vacuum when searching for target hairs, as there
remove hairs. filters can be individually is usually a lot of ‘background’ informa-
sealed. tion gathered in the form of debris and
material from the surface itself.
Shaking The object is shaken over a large collection Fabric type objects, e.g. pet rugs. Very quick method for May miss evidence that is stuck in the weave
funnel and any evidence collected in a retrieving loose of the fabric. Does not allow the exact
Petri dish. evidence. location of the evidence to be identified.
Scraping Use of a scalpel to remove very embedded Any surface which has hairs Enables quick retrieval of May damage hair during removal.
hairs. embedded in it, e.g. painted hairs from situations that
surfaces. other methods would not
be able to remove.
Combing Use of a seeded comb (a comb in which Pelage of an animal. Gently removes surface Sampling a moulting animal may cause
cotton wool has been pressed into the hairs without pulling out problems when trying to identify any
base of the teeth or brush which removes hairs from the animal. target hairs present on the comb.
and retains extraneous target hairs
(McKenna and Sherwin, 1975).
Filtering Uncommon technique involving the use of Samples which are heavily Allows large numbers of Care is required to ensure that the
different solvents to remove debris and contaminated with soil and other hairs to be quickly solvents do not alter the hair evidence
extract hairs from contaminated samples. debris, e.g. buried samples. extracted from soil. in any way.
 Forensic Examination of Animal Hair 67

6.4.2 Recovery of control aka known bags. If the hairs are wet with body fluids
hair samples that are to be analysed for DNA, then the
hairs should be packaged in a paper evi-
Control samples are hairs that have been dence bag and stored in a freezer.
taken as reference hairs from any animal
that could be involved with the crime or
could have transferred hairs to any other 6.4.4 Documentation of evidence
animal, object or person linked to the crime.
The taking of control samples from animals
Normal documentation of evidence applies
is usually undertaken after a police or court
with animal hair, including the need to
request. If possible, 20–30 hairs should be
photograph evidence in situ and fully docu-
taken from different points on the animal,
ment when, where and who obtained it,
ensuring that all hair types have been re-
along with a unique identifying number
trieved and the samples represent the differ-
(Lenertz, 2001). Additional information is
ent lengths and colours present on the pel-
required for animal hair evidence when re-
age. It has been suggested that a total of
trieving control hairs either for comparison
400–500 hairs should be retrieved for com-
to a questioned hair or for a reference col-
parison purposes (Suzanski, 1988). Control
lection. This may include the following:
samples from animals should ideally be
taken from a clean uncontaminated area • Known species and sub-species.
and depending on the purpose of the ana- • Gender.
lysis, for example, whether the hairs are for • Age (Wildman, 1961).
comparison only or for the analysis of the • Any known hair or skin diseases.
presence of drugs, additional consider- • Body area sampled (Wildman, 1961).
ations may be needed. Ideally hairs should • Method of hair removal.
be gently combed from the body, so as to
allow the full hair to be collected (Wildman,
1961), but if this is not possible, hairs can be
removed by cutting them close to the skin. 6.5 General Structure of Hair

Hairs consist of three regions; the root

(proximal end), the shaft, and the tip – also
6.4.3 Packaging and storage known as the shield region in animal hairs
(distal end).
Prior to packaging, certain anti-tampering In addition to this, hairs are composed
and anti-contamination procedures may be of three main types of cell: cuticle, cortical
required. If a tape lift has been created, the and medulla. If a hair is envisaged as a pen-
edges of tape need to be sealed with add- cil, the central graphite column of the pen-
itional tape. The evidence tape also must be cil is the medulla, the wood surrounding
scored with a scalpel so that it marks the the graphite is the cortex, and the paint on
acetate backing, but avoids damaging any the outside is the cuticle. Figure 6.1 dem-
evidence, protecting against fraudulent re- onstrates the basic structure of an animal
placement. The type of evidence bag used hair. Each area provides information about
for hair evidence depends on the police the hair and can contain distinguishing fea-
force and available equipment, but there are tures. The cuticle is made up of overlap-
recognized procedures for the packaging ping scales that usually do not contain pig-
and storing of such evidence. Loose hairs ment (Partin, 2003). The cortex, consisting
should be stored first in a labelled paper of spindle-shaped cells, makes up the main
wrap, sometimes referred to as ‘drug wraps’ component of the hair, contains pigment
and then placed in a plastic evidence bag. granules and may also include small irregular-
Tape lifts and vacuum filters should be fully shaped air spaces called cortical fusi and
sealed and also placed in plastic evidence larger solid oval structures called ovoid
68 C. Gwinnett

Inner cuticle margin Cortical fusi

Pigment granules
Cortex
Ovoid body
Cuticle

Proximal

Medulla

Distal Cuticle scales

Fig. 6.1 . Basic structure of an animal hair.

bodies. The medulla is a central core of used. A variety of questions beyond the
shrunken cells with the spaces between the species of the hair may be asked of the fo-
cells filled with air, and whose structure can rensic hair analyst regarding a case, either
vary dramatically in animal hair (Deedrick by the police or in court. These may in-
and Koch, 2004). clude the following:
• Do all species of animal have a com-
bination of unique hair characteris-
6.5.1 Types of hair tics?
• How confident are you that the hair has
The complete covering of hair over a mam- originated from the particular animal in
mal is called the pelage. There are five main question?
hair types that cover a mammal’s body, serv- • How and when did the hair evidence
ing particular functions such as heat preser- transfer to the scene/victim?
vation and sensory aids. Table 6.2 describes • Could the hair have been transferred in-
these five hairs and their general features. nocently?
These questions need further work beyond
the use of analytical techniques and need to
6.6 Forensic Animal
take into account processes such as transfer
Hair Analysis and persistence and commonality of different
characteristics, covered in Section 6.8 below.
Forensic animal hair analysis focuses most
commonly upon species identification, but
may also include comparisons between target
samples and controls, the latter being much 6.6.1 Stages of hair analysis
more difficult. In addition to this, analytical
techniques may be employed to ask specific Analysis of hair evidence usually starts
questions about the hair, e.g. about the presence with general observations about the num-
or absence of drugs. This chapter will concen- ber, condition and position of the hairs
trate on the microscopy of animal hair, as this is found. Analysis will also include some
the most versatile and common technique macroscopic observations, where the hairs
 Forensic Examination of Animal Hair 69

Table 6.2. Animal hair types.

Hair Type Found? General Characteristics Evidential Value

Vibrissae Nostrils and muzzle Generally coarser than other Low

(whiskers) hairs and are thickest at Limited variations in this hair type
the root end. between species mean limited
value for identification and
comparison (Teerink, 1991).
There are some exceptions,
such as tiger and leopard
vibrissae, which exhibit
differences in cross-sectional
shape (Partin, 2003).
Over-hairs Main pelage Longer than other hairs present on Low, limited value for
the pelage. Generally coarse, identification and comparison.
straight with elongated tips.
Under-hairs Main pelage Usually shorter and finer than Medium
the other hairs and show Can aid species identification, but
uniform thickness from the do not hold the same value as
root to tip ends. guard hairs.
Guard hairs Main pelage Commonly make the bulk of the High
pelage; coarser and longer Have the greatest significance in
than under-hairs but shorter species identification due to the
than over-hairs. interspecies variation present
and the most useful when
undertaking a comparison
between a control and target
hair (Suzanski, 1988).
Bristle Found on the body Generally thick hairs with forked Low
of animals such as tips and with either absent,
domestic and wild narrow or intruding medullas
pigs and boar (Deedrick and Koch, 2004).
Cross-sections are usually
oval, circular or oblong.

are placed upon a contrasting backing to the control and the target sample, to be
allow general features such as colour, viewed at the same time under the same
length, shaft profile and condition to be ob- conditions.
served and can allow samples to be divided Microscopy is debatably the most im-
into smaller groups, e.g. under hairs and portant stage of analysis and its advantages
guard hairs. Shaft profile can sometimes be include the following: being a non-destruc-
particularly useful when identifying spe- tive technique; relative speed (important for
cies; for example, deer hairs have a dis- timeliness of analysis, case throughput and if
tinctive crimped appearance (Deedrick and repeat measurements are needed); and its in-
Koch, 2004). expensiveness, after the initial outlay (very
The next stage of analysis is the use of few consumables). Sample preparation for
high-powered microscopes in the form of a fibres to be used for using brightfield micros-
compound microscope (for the use of copy is very simple, typically involving only
bright field microscopy) or a comparison placing the hair in a mounting medium be-
microscope. Comparison microscopes are tween a glass slide and cover slip.
particularly popular, as they comprise two In some situations a polarized light
high-powered microscopes connected by a microscope may be used; this allows the same
bridge which allows two samples, i.e. observations as a compound microscope, but
70 C. Gwinnett

also allows qualitative and quantitative meas- or Meltmount® (RI = 1.539) are required for
urements using plane polarized light and be- microscopical analysis. Refractive Index is
tween crossed polars (where the sample is the measure of the bending of light when
placed between two polaroid films). Although passing from one medium to another. Gener-
not regularly used by animal hair analysts, it ally, mounting mediums are thought to be
does provide additional information about the best when they have a similar RI to the hair
hairs’ optical properties, such as their interfer- (keratin RI = 1.548) so as to allow a clear
ence colours seen under crossed polars. This view of the internal characteristics of the
property has been used in the analysis of hair (Petraco, 1987). Wildman (1961) de-
exotic animal hairs (Partin, 2003). scribes the use of liquid paraffin (RI = 1.47)
Scanning Electron Microscopes (SEM) as a useful mountant, which is somewhat
utilize high-energy electrons to scan the lower in RI than keratin, but allows both the
surface of the hair and may be used after internal features and surface characteristics
compound microscopy to create high reso- to be viewed adequately. If the hair needs to
lution, three-dimensional images at very be removed from the microscope slide and
high magnifications, allowing characteristics mountant, then this is possible by cracking
such as the scale pattern to be more clearly the glass cover slip and applying a few drops
viewed, this can be seen in Bahuguna and of Tissue-Tek® Tissue-Clear®or xylene and
Mukherjee’s (2000) work on identifying then gently removing the loosened hair.
Tibetan antelope hairs.
Finally, further analytical techniques 6.6.2.2 Scale casts and impressions of
may be utilized if additional information the animal hair surface
is required, such as dye analysis or drug
Prior to mounting on a microscope slide, a
analysis. Common techniques are High Per-
scale cast (or cuticle slide) can be produced
formance Liquid Chromatography (HPLC)
to allow the outer scale profile to be deter-
and Thin Layer Chromatography (TLC) for
mined. To do this, a thin layer of gelatine
extracted dyes, and radioimmunoassays (RIA)
(10%–20%) (Teerink, 1991), polyvinyl acet-
and chromatography techniques, such as Gas
ate, Meltmount® (Petraco, 1987), or clear nail
Chromatography (GC), for the analysis of ex-
polish can be painted on to a microscope
tracted drugs (Gratacos-Cubarsi et al., 2006).
slide in a uniform thickness. The hair sam-
ple can then be placed gently into the sub-
strate, leaving the end of the hair out of cast-
6.6.2 Microscopy preparation
ing material for easier removal, and allowed
of animal hairs
to dry. If PVA is used, an additional slide can
be placed on top of the hair and the slide
Any target samples, or even control samples, gently heated to the melting point of the PVA
which are covered in debris or body fluid must and then allowed to cool before removal of
be cleaned prior to analysis. This can be com- the hair (Wildman, 1961). The hair can be
pleted by gently washing with distilled water rolled if a full impression is required as de-
and a mild detergent if necessary. Organic scribed by Wildman (1954), although this
solvents such as isopropyl alcohol or acetone may damage the shaft and is not always ap-
can be used to remove grease and other sur- propriate, as the shield of the animal hair is
face impurities (Ogle and Mitosinka, 1973). normally slightly flattened in cross-section
If DNA analysis is possible, hairs (Huffman and Wallace, 2012). When the sub-
should initially be mounted in distilled water strate is fully dry, the hair can be carefully
so as to prevent any nuclear DNA being des- removed and the resultant cast can be viewed
troyed by a mounting medium. under a high-power light microscope.

6.6.2.1 Creating a whole mount 6.6.2.3 Medulla slides

Mounting mediums such as Entellan® (Refra- When the formation of medulla is difficult to
ctive Index (RI) = 1.49–1.51), DPX (RI = 1.52) visualize, a medulla slide may be produced,
 Forensic Examination of Animal Hair 71

which removes the air present in the me- and interpretation of the evidence. In an in-
dulla to allow for a detailed view of the vestigation, an analyst may be expected to
structure. This is achieved by infiltrating the analyse a few hairs, partial or complete hides,
medulla with xylene or paraffin oil, which an object from a crime scene or a finished
makes it transparent when viewed under a product such as a fur coat or hat (Linacre,
microscope (Teerink, 1991; Linacre, 2009). 2009); therefore analysis methods must be
To do this, the hair is cut at various positions adaptable but still recorded in a robust and
using a razor blade, such that the xylene can reliable manner.
seep into the medulla. This process can take It is also beneficial to provide sketches
up to three hours. To make these permanent, and/or photomicrographs of the hairs.
the oil can be replaced with a medium such Sketches should include all three areas of
as Canada balsam (Teerink, 1991). The pro- the hair (root, shaft and tip) and be large, in
duction of these slides is particularly useful ink, annotated, and signed and dated.
for lightly pigmented hairs, but with some It is logical for an analyst to start at the
highly pigmented hairs, such as some pri- root end, move through to the shaft area
mates and black bears, there must be add- and then to the tip to identify any variation
itional treatment of the cortex to make it in characteristics and to view the whole
transparent. This can be achieved by sub- hair. Each area can provide information
merging dry hairs in hydrogen peroxide and specific to that region; for example, the
a few drops of ammonia solution until the root can provide information about the
desired lightness is achieved (Linacre, 2009). growth stage of the hair, the shaft can pro-
vide information about the pigment and
medulla and the tip can provide informa-
6.6.3 Microscopical analysis tion about damage and whether a fork or
of animal hairs split is present.
For each of the cortex, medulla and cu-
In microscopical analysis of hairs, a balance ticle regions, there are particular observa-
exists between observing the whole hair to tions that are deemed useful for the analysis
identify species and/or any similarities and of hairs; these are listed in Table 6.3.
differences between control and target hairs. Figures 6.2 and 6.3 demonstrate ex-
To achieve this, a systematic approach is re- amples of standard animal hair analysis
quired. Analysis evidence sheets can be used forms and the categories used to describe
that provide a systematic method for noting the observations stated in Table 6.3. Further
down relevant characteristics, sketches and descriptions of the key observations can be
comments using standardized terminology; found in Section 6.7, ‘Species Identification
these analysis sheets simplify comparison from Animal Hair’, below.

Table 6.3. Microscopical observations for the cortex, medulla and cuticle.

Hair Region Characteristic

Cortex Commonly noted: colour/hue, pigment density, pigment distribution, presence/absence of

ovoid bodies.
Additional observations: pigment granule shape, pigment granule size, texture of cortex,
presence/absence of cortical fusi.
Medulla Commonly noted: medulla distribution/type, medulla opacity, Medullary Index (MI)
(Medulla diameter/shaft diameter).
Additional observations: form of the medulla margins (straight, fringed, scalloped)
(Teerink, 1991).
Cuticle Commonly noted: cuticle thickness, scale pattern, scale position in relation to longitudinal
direction of the hair, scale edge shape, distance between scale edges.
Additional observations: distinctness of the inner cuticle margin.
72 C. Gwinnett

ANIMAL HAIR EXAMINATION – SHEET 1

Case reference:
Page——of
Macroscopic characteristics
Hair
evidence
number
Length
(cm)

Shaft
profile

Colour

Presence
of banding
General description:

Examined by:

Notes by: Day: Date: Time:

Fig. 6.2. Example of a standard animal hair form: Part 1 (macroscopic observations and sketches).

In addition to these observations, the root diameter of the shaft should be measured
growth stage may be noted: this can be cat- in micrometres using a calibrated eye-piece
egorized into anagen (active growth stage scale, and variation along the length of the
with the presence of nuclear material), cata- hair should be noted.
gen (transitional growth stage with limited Depending upon the particular crime,
nuclear material) or telogen (dormant stage and when and where the hair was found,
where hairs are readily shed and no nuclear animal hair evidence may have been subject
material present) (Robertson, 1999). The to external influences such as weathering
 Forensic Examination of Animal Hair 73

ANIMAL HAIR EXAMINATION – SHEET 2

Case reference:
Page——of
Microscopic characteristics
Hair evidence number
Pigment None
density Light
Medium
Heavy
Pigment Even
distribution Central
Peripheral
One-sided
Medulla None
distribution/ Broken
type (fragmented/interrupted)
Unbroken/continuous
(lattice/aeriform
lattice/simple/vacuolated)
Ladder
(uniserial/multiserial)
Miscellaneous
(globular/stellate/intruding)
Scale edges Smooth
shape Crenate
Rippled
Scalloped
Dentate
Distance Close
between Near
scales Distant
Scale pattern Mosaic (regular/irregular)
Wave
(regular/irregular/single
chevron/double
chevron/streaked)
Petal
(broad/elongate/diamond*)
Transitional

Ovoid bodies (Y/N)

Shaft diameter (μm)
Root shape
Tip shape
Medullary Index (MI) = Medulla
diameter/shaft diameter
Other
Examined by:
Notes by: Day: Date: Time:
* note whether narrow or broad diamond

Fig. 6.3. Example of a standard animal hair form: Part 2 (microscopic observations).

(Chang, 2005), causing change in the mor- cigarette burns, irons or complete burning
phological features, but nevertheless pro- of the hair with accelerants. Hair samples
viding additional information about the found in bedding, discarded collars and at
case. In certain cruelty cases, animals may crime scenes can indicate the temperatures
have been exposed to heat sources such as that hair has been exposed to.
74 C. Gwinnett

When hairs are exposed to heat, changes related species can show similar characteris-
in colour, swelling and bubbling of the hair tics (Wildman, 1961).
may occur. Research conducted by Pangerl For species and sub-species identifica-
and Igowsky (2007) on human hairs indicated tion, there are five main characteristics:
that variables such as temperature, exposure scale morphology; medulla type; medullary
time, and how the heat is applied to the hair fraction (MF); colour banding; and root shape.
must be considered to fully interpret this The nomenclature for the different charac-
type of damage. Research conducted by Ay- teristics differs between ID keys and guides,
res (1985) identified that colour changes oc- but the following categories for each charac-
curred in hairs when exposed to a hot plate, teristic primarily combines the terminology
but when exposed directly to flame, colour and species examples used by Appleyard
changes were absent; however, the presence (1960), Wildman (1961), Petraco (1987),
of charring and bubbling was observed. Moore (1988), Teerink (1991), Partin (2003),
Work conducted at Staffordshire University Deedrick and Koch (2004) and Linacre (2009).
has shown that it is also possible to identify All examples are for guard hairs unless other-
the presence of accelerants on even a few wise stated.
strands of hairs that have been in close range
of an accelerated fire, using headspace gas
chromatography.
Other environmental factors that cause 6.7.1 Scale morphology
damage to the hair, such as crushing, insect
damage and fungal damage, can alter the ap- Broadly the scale pattern of animal hairs
pearance of the hair, but also potentially pro- can be classified into two main groups: cor-
vides an evidentially useful characteristic. onal (where the scales go around the entire
For example, hair from a decomposing body shaft, completely encircling it); and imbri-
may exhibit a decomposition band, i.e. the cate (where there are multiple scales encirc-
section of hair lying below the skin surface ling the shaft). Further classifications can be
has darkened in colour due to the decompos- made of the scales on the cuticle by observ-
ition process (Linch and Prahlow, 2001). ing four main characteristics.

1. Scale position in relation to the longitu-

dinal axis: this can be categorized as trans-
6.7 Species Identification versal (where the scales are at right angles to
from Animal Hair the longitudinal axis and appear to have a
greater width than length – seen in Fig. 6.4f);
It has been noted by Petraco (1987) and longitudinal (where the scales are aligned
Moore (1988), that with use of their schemes with the length of the hair and are longer
of identification, even inexperienced exam- than they are wide – seen in Fig. 6.4b); or
iners can accurately identify the species of intermediate (where the width of the scales is
an animal from its hair, albeit with use of the same as the length – seen in Fig. 6.4a).
other resources, such as reference materials 2. Shape of the scale margin: this is the
(Moore, 1988). This comment should be taken shape of the distal end of the scale, which
with caution, as it is recognized that animal can be smooth, crenate (shallow and rela-
hair identification is one of the more difficult tively pointed indentations), rippled, frilled,
analyses attempted by forensic scientists scalloped or dentate (pointed, like teeth).
(Wildman, 1961; Moore 1988). Possible ex- 3. Distance between the external margins of
planations for this include: the variation that can the scales: this is usually categorized as close
exist within a species (Moore, 1988); the vari- (as seen in Fig. 6.4g); near (as seen in Fig. 6.4a);
ation in terminology used in species identifica- or distant (as seen in Fig. 6.4b). Sometimes
tion keys; the subjective nature of the analysis (no scale count is also completed, which counts
single characteristic will allow identification of a the number of scales per unit of measure,
species); and the fact that hairs from closely e.g. 40μm (Rosen, 1974).
 Forensic Examination of Animal Hair 75

(a) (b) (c)

Broad petal – Seal Elongate petal Broad diamond

aka pectinate – Otter petal – Fox,
Badger, Dog
(d) (e) (f)

Narrow diamond Mosaic Regular wave – Bear,

petal – Harvest Mouse Regular – Cat, Dog, Beaver, Bison
European Hedgehog
Irregular – Lion, Cat

(g) (h) (i)

Irregular wave – Orangutan, Streaked – Bank Single chevron – Daubenton’s

Aardvark Vole Bat

(j)

Double chevron – Squirrel,

Rabbit

Fig. 6.4. Animal hair scale patterns.

4. Scale pattern: this describes the overall of animals that exhibit these. In addition to
shape and regularity of the outer scales. Fig- these, a pattern may also be transitional,
ure 6.4 demonstrates the most common pat- which is the presence of more than one pat-
terns seen in animal hair, with some examples tern along the length of the hair.
76 C. Gwinnett

6.7.2 Medulla types further categorized by their structure (and

width, for certain types). Teerink (1991)
The medulla can vary dramatically between also further categorized medulla by the form
species and even subtle differences can be of the medulla margin, i.e. the silhouette of
seen between sub-species. First, medullas the medulla’s edges, as straight, fringed or
can be categorized by their distribution as scalloped. Figure 6.5 demonstrates the most
being unbroken, broken (e.g. interrupted common medulla structures seen in animal
and fragmented which is also seen in human hair, with some examples of animals that
hair), laddered, or miscellaneous; and then exhibit these.

Unbroken
(a) (b) (c)

Amorphous/simple – Lion, Vacuolated/cellular – Red Lattice

Raccoon Fox Wide – Deer, Otter
Fine – Mink, Beaver

Laddered
(d) (e) (f)

Aeriform lattice – Muskrat, Uniserial – Rabbit Multiserial – Rabbit

Chinchilla, Opossum, (under-hairs) (guard hairs)
European Hamster

Miscellaneous
(g) (h) (i)

Stellate Globular – Seal Intruding – Horse

tail hair

Fig. 6.5. Animal hair medulla distributions and structures.

 Forensic Examination of Animal Hair 77

6.7.3 Medullary fraction (MF) complete guard hair. Moore’s (1988) scheme
aka medullary index (MI) mainly focuses upon animal species com-
monly found in the UK, but also incorporates
The medullary fraction (MF) is the ratio be- other species such as camel and llama. A com-
tween the width of the hair and width of the prehensive atlas of west-European hairs
medulla. The width of both the shaft of the developed by Teerink (1991) provides illus-
hair and the medulla are measured in micro- trations and photographs of cross-sections,
metres and can be compared as a quantita- scale casts, medulla slides and mounted sam-
tive characteristic or used to aid identifica- ples of guard and under-hairs from a vast
tion; for example, Peabody et al. (1983) range of animals. Smaller research projects
determined that the medullary fraction analysing particular species or geographical
could be used to reliably distinguish be- area either for casework or other environ-
tween dogs and cats. mental or scientific purpose are also very use-
ful to the hair analyst. Examples of these
types of study include Williams’ (1938) ID of
6.7.4 Colour banding mole and shrew hairs; Hilton and Kutscha’s
(1978) ID of coyote, dog, red fox and bobcat
Pigment distribution in animal hair may not hairs in Maine; Vineis et al.’s (2008) ID of
only differ across the width of the hair, but wild goat and domestic goat hair; and Mayer’s
also quite dramatically along its length. The (1952) examination of Californian mammals.
length, order, colour and number of bands In addition to these works, there are very use-
can help identify different species. For ex- ful online resources that present photomicro-
ample, badger hair can be differentiated graphs of different animal hairs, including
from dog and fox hair primarily by its dis- Deedrick and Koch (2004).
tinct white proximal end, black shaft and These guides are very important for fo-
white tip (Moore, 1988). rensic analysts who may not have come
across certain animal hair types in casework
previously. Some of these keys state differ-
6.7.5 Root shape ent characteristics for the same species type
but this is to be expected, as different
Further to identifying growth stage, animal sub-species will have been sampled for the
hair root bulbs can have particular shapes production of the keys and therefore the use
that are useful in identifying species. Ex- of multiple keys, to identify any variation
amples of this include deer (wine glass), and differences in interpretation, is advised.
horse (elongated), cow (elongated but with The breadth of animal hairs and their
a medulla present in the root portion), dog microscopical characteristics is huge and it
(spade) and cat (‘paint brush’ with the inclu- is advisable for a forensic analyst to have ref-
sion of fibrils) (Moore, 1988; Linacre, 2009). erence samples of a large range of animals for
comparison (Wildman, 1961). Samples may
be obtained from casework, museums, zoos
6.7.6 Species identification aids or commercially produced collections, such
the Arbidar Animal Hair and Fur Collection.
Research into the variation of morphological
characteristics in animal hair, comparisons of
different species and sub-species, and the de-
velopment of identification keys, reference
6.8 Interpretation of Animal Hair
collections and interpretation aids are abun- in Casework
dant. Examples of specific ID schemes in-
clude the following: Stains (1958); Appleyard 6.8.1 Conclusions from comparing control
(1960); Moore (1988); and Petraco (1987). and target hairs
Petraco’s (1987) scheme includes 25 genera
and was designed to allow quick and effective The interpretation of the characteristics
identification of these genera using only one observed between target hairs and control