鑑識科学，7(2)，95―111(2003) 95

―Review―

Forensic Examination of Soil Evidence

Yoshiteru Marumo

National Research Institute of Police Science

631, Kashiwanoha, Kashiwa, Chiba 2770882, Japan

(Received 18 November 2002: accepted 25 November 2002)

Soil can provide important information to criminal investigations as transfer

evidence because many criminal cases take place under circumstances such that soil
transfers to a criminal or victim. The variation in soils from place to place makes soil
valuable evidence to prove linkage between a suspect and a crime scene. Soil is a
complex mixture with a variety of mineralogical, chemical, biological, and physical
properties. Considering such complexity, a variety of methods have been developed
for forensic science purposes. Because minerals are an important component of
soils, mineralogical examination is essential in forensic soil identiˆcation.
Additionally, many other methods can be applied to raise the discriminating power,
but not all kind of methods need to be used. What is important is that examiners
select an appropriate combination of methods by considering the context of the soil
samples. This report summarizes a wide range of reports on the analysis of soil
components and of closely related materials such as plant fragments, pollen and
spores, and diatoms, with emphasis on the importance of screening tests consisting
of several simple techniques. The soil formation process involves parent materials,
temperature, water condition, vegetation, time, and the chemical processes of
solution, oxidation, reduction, and even human activities. The history of a soil's
development as the results of such complex soil formation process is strongly
re‰ected in soil color. The systematic observation of multiple soil colors is especially
useful for screening.

Key words : Forensic science, Soil identiˆcation, Soil color, Particle size
distribution, Mineral, Botanic material

1. Introduction Gross of Austria wrote, in his handbook on

Because of its ubiquitous nature as the criminalistics, that dirt on shoes could probably
surface of the earth, soil plays an important role can yield useful information for a criminal
as a type of transfer evidence that can establish investigation, but he did not include any actual
linkage between a crime scene and a suspect. case study or practical technology. According to
More than a hundred years ago, such a role for a report by Murray1), Georg Popp of Germany
soil evidence in a criminal investigation was ˆrst might have been the ˆrst scientist who actually
described. But it was in a ˆctional story of the examined soil evidence, which led to the solution
Sherlock Holmes series written by Sir Arthur of a murder case in 1904.
Conan Doyle. At almost the same time, Hans The evidential value of soil rests on the large
96 Yoshiteru Marumo

variation in its characteristics from place to was published in 1975. It has since been revised
place. Soil has extreme complexity, not only in to include recent technological developments
its components, such as minerals, oxides, and has the title Forensic Geology3).
organic matter, microorganisms, and parent Several handbooks and review papers have
materials, but also in its physical nature, such as addressed the complexity and diversity of
particle size and density. Consider, for instance, techniques available for forensic soil examina-
soils with granite rock as their sole parent tion48). The review paper by Demmelmeyer and
material. An almost unlimited number of kinds Adam7) discuss 178 reference reports that deal
of such soils are easily recognizable by with (1) visual comparison of morphology, (2)
diŠerences in color, mineralogy, and texture. preliminary examination with chemical reagents,
Soil diversity is the result of soil formation (3) particle size distribution, (4) density gradient
processes acting on a variety of parent materials. distribution, (5) polarized light microscopy, (6)
These processes strongly depend on topography, further physico-chemical analysis, (7)
climate, time, botanical and microbiological biochemical components, and (8) vegetable trace
activities, water conditions, and even human materials. An overview paper9) presented at the
activities. Moreover, there are millions of fossils, INTERPOL Forensic Science Symposium,
external matter such as pollen and spores, and which are held triennially, covers reports on
even artiˆcial materials that also contribute to technical developments of the previous three
the soil. Forensic soil examination is considered years.
very complicated because of the complexity of
soil, but such diversity allows us to identify soil 3. Collection of Soil Samples
samples with high discriminating power. The signiˆcance of soil identiˆcation largely
Because of the uniqueness of the forensic depends on the quality of sample collection;
purpose and the complexity of soil, a great many especially, the collection of control samples is
methods for forensic soil examination have been very important. The improper collection of
proposed and discussed. Successful soil control soil samples will cause an incorrect
identiˆcation depends on examiners selecting a judgment. Murray10) reported his experience
combination of methods suitable for the speciˆc with 30 soil samples that, even though they had
conditions of the soil evidence. The major object been collected systematically, were useless
of this paper is to present an overview of forensic because they did not represent the exact original
soil identiˆcation by summarizing the literature location of the questioned soil sample.
on the topic for the last three decades and to help Because of the heterogeneous nature of soil,
forensic scientists select the best combination of the signiˆcance of a match between questioned
methods appropriate for each case at hand. As and control samples must be interpreted taking
soil particles are one of the major components of into account the intra-sample variations in the
airborne dust, they frequently can be transferred questioned samples and that in the control
if a suspect touched a dusty surface such as a samples. Dudley and Smalldon11) suggested
door, windowsill, etc. This report, therefore, multiple sampling of control samples on the
includes a discussion of dust and other earth- basis of their studies of simulated crime scenes.
related materials such as plant fragments, Lenth12) and Evett13,14) recommend the use of
diatoms, pollen, and spores. probability theory to evaluate soil evidence.
Munroe15) has presented an excellent guide
2. Textbook and Review Papers to sampling for forensic soil examination written
The monumental work in forensic soil from the pedological viewpoint that soil varies
investigation is the textbook entitled Forensic both in a horizontal and vertical direction.
Geology: Earth Science and Criminal Control soil samples should be collected after
Investigation by Murray and Tedrow2), which careful visual inspection of soil color and texture
 Forensic Examination of Soil Evidence 97

changes, both horizontal and vertical, and soil played an important role in the criminal
samples of diŠerent colors or textures should not investigation1,17).
be mixed even if they were taken from sites in A police o‹cer who happened to look at a
close proximity. The soil sample should be kept man arrested for a minor crime said ``That's the
unbroken as much as possible to preserve its worst case of dandruŠ I have ever seen.'' Later,
structure. There is also a detailed description of it proved to be not dandruŠ but diatomaceous
sampling from a vehicle. earth, and it compared species-to-species with
the insulation material of a safe that had been
4. Macroscopic and Low-Power ripped open on the previous day1,10).
Microscopic Observation In other murder cases, shell fragments in
Macroscopic and low-power microscopic sand, lava fragments with distinctive features,
observations are an important initial step in and augite crystals with inclusions were key
forensic soil comparison. DiŠerences in the evidence in the criminal investigations.
visual features between questioned and control
samples mean that they originated from diŠerent 5. Screening of Soil Samples
locations, suggesting that any further The signiˆcance of a match between
examination is not necessary. If unique foreign questioned samples and control samples must be
particles, soil aggregates, or other trace evidence interpreted in light of the variations among the
are found in soil samples, these particles should questioned samples and that among the control
be removed and preserved separately for further samples. The number of soil samples collected
examination16). With luck, those unique particles for comparison, therefore, must inevitably
may be found in both a questioned soil sample increase to obtain interpretations with evidential
and a control one. Further examination can value. Simple and rapid screening methods are
focus on these unusual materials in the soil, required for successful identiˆcation of a large
which at this step is regarded as only background number of soil samples. Color comparison and
of evidence. the determination of particle size distribution of
Some interesting cases in which unique soil samples can be carried out quite easily, and
particles were the key to solving a crime have the combined data from those measurements can
been reported. For example, in a murder case in be quite useful for discriminating among
California, the victim's body was abandoned at samples.
an oil-well apron, where gravel that had been 5.1 Soil Color
transported from 300 miles further south had Soil color, which re‰ects the pedogenic
been used. In the soil evidence collected from a environment and history18), is one of the most
suspect's car, rock fragments were found and distinguishable characteristics of soil. Generally,
identiˆed as having the same origin as those the Munsell Color System is used for soil color
around the oil-well apron1,10). determination, and the color is expressed by the
Blue thread was decisive for solving a rape combination of hue, value, and chroma. Soil
case in upper Michigan. Three ‰owerpots had Color Standards19,20), in which color chips from
been tipped over and the soil spilled on the ‰oor the Munsell Color System that cover the typical
in the struggle. Potting soil on the suspect's shoe soil-color range are arranged, are widely used in
was compared with that spilled from those the laboratory and the ˆeld.
‰owerpots. Small clippings of blue thread were Dudley21) examined the usefulness of color
found both in the ‰ower pot sample and on the examination for forensic soil discrimination, and
shoe of the suspect1). proposed multiple color comparisons consisting
In a rape case in New Jersey, the rape took of three color measurements for each soil
place in a vacant lot that had been used for a coal sample, carried out on air-dried, moistened, and
pile 60 years previously, and coal fragments ashed (850° C, 30 min) sample. He concluded that
98 Yoshiteru Marumo

ashing had very good discriminatory potential, important procedures when screening soil
but moistening was less useful. Moistened soil samples. The method generally used to separate
color, however, should always be borne in mind, particle size fractions is sieving, and the relative
particularly when dry and ashed colors oŠer no abundance of particles in diŠerent size ranges,
possibility for discrimination. the so-called particle size distribution, is useful
Antoci and Petraco22) proposed the use of to compare soil samples. Some of the factors
specially prepared paperboard for side-by-side that can aŠect the suitability of dry sieving for
comparisons of a soil sample directly with the the determination of particle size distributions
standard color chips, and Janssen et al.23) were studied by Wanogho et al.28). Dried soil
proposed that color measurements should be samples, after decomposition of organic matter
carried out on the clay fraction in order to by hydrogen peroxide, were subjected to dry
minimize variation caused by heterogeneity of sieving with a sieve shaker. The results indicated
the soil particles. that the larger the mass of the sample used, the
Color comparison of air-dried soil alone is better the degree of reproducibility. The most
insu‹cient for sample discrimination. Soil color reasonable sample weight to adopt for analysis
can be seen as a result of a mixture of soil was 1 g or more. They concluded that the dry
materials. Iron oxides give soil a reddish, sieving technique was not preferred over wet
brownish, or yellowish color, and humic sieving29). Factors aŠecting the suitability of wet
substances make the soil darker. The parent sieving with a shaker, in comparison with dry
materials also in‰uence the soil color. Sugita and sieving, were also studied by Wanogho et al.30)
Marumo24) studied the usefulness of soil color and Robertson et al.31) The results by wet sieving
for forensic examination in light of these showed less variation than those by dry sieving,
pedogenic soil properties, and proposed a multi- but they concluded that simpler wet-sieving
step soil color comparison method, in which procedures were necessary because those using a
successive color determinations are made after shaker were rather complicated and required a
air drying, moistening, decomposing organic large amount of sample.
matter, removing iron oxide, and ashing (850° C, Measurement of the particle size
30 min). This method was applied to 73 soil distribution of the ˆner particle size fractions of
samples collected systematically with reference soils was studied by Dudley32), using a Coulter
to a geologic map and a land-use map25). In Counter, and by Wanogho et al.33), using a laser
contrast to the results of Dudley21), color after diŠraction technique.
ashing did not raise the discriminating power, Dry sieving is easy to perform, but its
and moistening also did not allow eŠective reproducibility is poor if many aggregates are
discrimination among dark-colored soil samples present in the soil samples. Clay, especially,
with a value of less than 3 in the Munsell system. strongly coagulates soil particles, therefore, dry
The combination of soil colors determined after sieving cannot be applied to the ˆne particle
air drying, decomposition of organic matter, and fraction. Wet sieving is required for fractions
removal of iron oxides enabled up to 98 of the less than 0.05 mm in size. A study by Wanogho
soil samples to be discriminated, compared with et al.34) indicated that wet sieving provides good
84 by the single comparison of air-dried soil reproducibility.
color. For comparison of color after Marumo and Sugita35,36) examined the
decomposition of organic matter and removal of validity of particle size distribution for forensic
iron oxides, it is preferable to use clay fractions soil discrimination. As far as reproducibility is
because of their homogeneity26,27). concerned, wet sieving is preferable. But to
5.2 Particle Size Distribution simplify the procedure, wet sieving should be
Comparison of particle size, which is an employed only for the separation of the ˆne (＜
apparent visible feature, is one of the more 0.05 mm particle size) fraction, and dry sieving
 Forensic Examination of Soil Evidence 99

can be applied to the coarser particle fraction. In 7. Rock Fragments and Sand Particles
order to obtain better reproducibility for small As the brownish color of soil mineral
soil samples, they recommended that only three particles are mostly the result of staining by iron
particle size classes should be used: ＜0.05, 0.05 oxides, removal of iron oxides is required prior
0.2 and 0.2 2 mm. The results of the to mineralogical examination. Generally,
examination of 73 soil samples collected Aguilera and Jackson's method43), in which
systematically showed that 88 of samples sodium dithionite is used to reduce the iron
could be discriminated by this method. oxides, is applied to the coarser particle size
fractions of the soil. The 0.22 and the 0.050.2
6. Density Gradient Distribution mm fractions are examined for rock fragments
The observation of distribution of soil and sand particles, respectively.
particles in a density gradient column is the most 7.1 Rock Fragments
traditional method used for forensic soil Rock fragments can provide useful
comparison. At present, however, only a few information not only for sample-to-sample
forensic science laboratories are using this identiˆcation but also for predicting the original
method. site of soil evidence, because rock fragments
A density gradient tube is prepared by strongly re‰ect the geological characteristics of
layering liquids of varying densities in a glass the soil. Rock fragments can be easily examined
tube such that they gradually become less dense under a low-power binocular microscope
from the bottom to the top. Liquids of varying without any sophisticated skill. The importance
densities are made by mixing heavy and light of this coarse soil fraction was thoroughly
liquids in various proportions. Generally, studied by Ugolini et al.44), who asserted that
bromoform (2.89 g/mL) or tetrabromoethane distinct mineralogical diŠerences from site to site
(2.96 g/mL) is used as the heavy liquid, and provide useful information to criminal
bromobenzene (1.50 g/mL) or ethanol (0.789 investigations.
g/mL) is used as the light liquid. A soil sample Soil evidence collected from a car ‰oor is
that has been dried and gently crushed is placed the most di‹cult evidence to evaluate by forensic
onto the density gradient tube37). science, because many diŠerent materials from
Many studies have been carried out with the diŠerent sites are brought into the car as the
intention of improving the reproducibility of the driver repeatedly gets into the vehicle.
density gradient tube method3840). However, Nevertheless, if examiners discover rock
careful studies by Chaperlin and Howarth41) fragments with distinctive features, these
revealed that the comparison of density gradient fragments can provide very important
patterns is very dependent on the observer, so information to the criminal investigation.
reproducibility can be a problem. Murray and Rapp45) reported an interesting case in which
Tedrow3) stated that the density gradient method small gravel found on the ‰oor of the suspect's
has severe limitations, and little weight should be car played an important role in the criminal
given to the results of this method. investigation.
Petraco and Kubic42) recently expanded the 7.2 Sand Particles
density gradient range from 2.891.50 g/mL to Microscopic examination of sand particles
4.241.00 g/mL by using Clerici's solution (CS, is an essential technique for forensic soil
saturated solution of thallium malonate- comparison because the sand fraction is one of
formate) and distilled water (DS) instead of the core components of soil, and mineralogical
bromoform/bromobenzene or bromoform/ information on sand particles can be used to
ethanol. This density gradient column (CS/DS) infer reasonable relationships between
can cover the whole density range of soil mineral questioned soil samples and control ones. As
particles. examination of optical properties using a
100 Yoshiteru Marumo

polarizing microscope requires experience and would be very useful for forensic soil
skill, as well as long and tedious eŠort, recent examination.
studies for forensic purposes have tended to Application of a scanning electron
focus on simpliˆcation of the method. Heavy microscope equipped with an energy dispersive
mineral composition is informative in forensic xray spectrometer (SEMEDX) to mineralogi-
soil comparisons, but, as heavy minerals are not cal examination is the most promising approach
as abundant in soil, heavy minerals and light for the future. McVicar and Graves51) developed
minerals should be subjected to mineralogical a particle search and analysis technique using an
examination separately. automated SEMEDX system. Sand particles
Graves46) has proposed a conventional were identiˆed according to mineral
microscopic method for forensic scientists to classiˆcation rules that had been formulated
determine soil mineralogical composition. The beforehand on the basis of the analysis of known
classiˆcation of minerals into 38 categories is minerals.
carried out by sequentially changing 14 optical Park et al.52) attempted to apply image
conditions. Junger47) used an even more processing to pictures taken of sand particles
simpliˆed classiˆcation system with 25 under a polarized microscope. The color
mineralogical categories. We use 15 categories: distribution calculated from the image
quartz, alkaline feldspars, plagioclase, volcanic processing was compared between soil samples.
glass, biotite, hornblende, hypersthene, augite, The results of this method do not completely
olivine, chlorite, zircon, magnetite, rock correspond to the mineralogical composition,
fragments, weathered particles, and other. Of but it can be useful for sample-to-sample
course, mineralogical categories should be comparison.
changed according to local variations and the The examination of sand particle
speciˆc geological and pedogenic situation. If morphology for forensic science purposes has
unique minerals other than those categorized been reviewed by Fitzpatrick and Thornton53).
minerals exist in the soil samples, they should be
recorded. 8. Clay Minerals
Xray diŠractometry (XRD) can also be Since clay minerals are secondarily
applied to the identiˆcation of primary minerals. produced in a soil pedon from parent materials
The amount of sample required for XRD can be during the soil-forming process, clay
reduced to 2 mg by using a quartz nonre‰ecting mineralogical composition re‰ects both the
sample plate48). However, it should be noted that pedogenic environment and the parent material,
XRD cannot detect minor minerals composing such as sediments or metamorphic rocks. As clay
less than 5 of the sample. minerals are too small for optical microscopy to
A staining method has been reported that be applied, identiˆcation of clay minerals relies
simpliˆes light mineral identiˆcation49). on instrumental analyses. Marumo et al.5458)
Sequential treatment with potassium chloride, studied clay mineralogical analysis from the
sodium cobaltinitrite, and hematein stains viewpoint of forensic science using XRD,
alkaline feldspars yellow and plagioclase purple, infrared absorption spectrometry (IR), and
whereas quartz remains unstained. diŠerential thermal analysis (DTA). The particle
An attempt has been made to use size fraction less than 0.05 mm, after
identiˆcation charts for mineralogical decomposition of organic matter and removal of
examination with a polarized microscope. iron oxides, was subjected to the instrumental
Ishiwatari50) proposed a useful identiˆcation analyses. As the crystals of most clay minerals
chart including 200 minerals for identiˆcation of have a platy structure, the basal spacings are
minerals in rock specimens. If this identiˆcation recorded by XRD. For each soil sample, two
chart were simpliˆed for use with soil minerals, it kinds of oriented specimens were prepared from
 Forensic Examination of Soil Evidence 101

magnesium-saturated (Mgclay) and potassium- Humic substances are a group of vastly

saturated (Kclay) clay. XRD was carried out on complicated organic substances that are
the Mgclay specimen successively after air produced by biological decomposition of plant
drying and after solvation with a glycerol bodies and other biotic materials. A variety of
solution, and on the K clay specimen analytical methods for humic substances have
successively after air drying, after heating at 300 been proposed for soil science investigations, but
°C, and after heating at 600° C. IR was all those methods are very complicated and time-
successfully applied to clay specimens for the consuming, and they require more than several
identiˆcation of gibbsite and kaolinite minerals. grams of soil sample. Honna et al.61) developed a
DTA was useful for identifying allophane, which simple technique to evaluate humic substances,
cannot be identiˆed by XRD or IR. The validity in which a soil sample (0.51 g) extracted with
of the clay mineralogical analysis for forensic 0.5  sodium hydroxide is subjected to
soil comparison was conˆrmed by applying these absorbance measurements.
methods to soil samples collected from a region The chromatographic pattern is a
where all soils had developed from granite. As a convenient method for comparing organic
result, the clay mineral composition showed matter between soil samples. Siegel and
variation re‰ecting topography57). Precord62) and Reuland et al.63) proposed the
McAlister and Smith59) proposed a rapid application of high performance liquid
technique for clay specimen preparation for chromatography (HPLC) with reverse phase
XRD analysis whereby clay suspensions are columns to soil extracted with organic solvents
precipitated onto membrane ˆlter discs by such as methanol or acetonitrile. Nakayama et
syringe injection under pressure. al.64) applied pyrolysis gas chromatography
(PyGC) to 113 soil samples collected from
9. Organic Matter Tokushima prefecture, and used mass
The soil ecosystem contributes greatly to spectrometry (MS) to further identify major
soil development, resulting in extreme soil peaks on the pyrogram. The pyrogram patterns
diversity. There is incredible complexity in the showed good reproducibility within the same
soil ecosystem involving soil water, the cation- soil, and large variation among soil samples of
exchange and adsorption capacity of clay diŠerent origin.
minerals, temperature, pH, soil atmosphere, Cox et al.65) recently developed an excellent
solar radiation, oxidation-reduction potential, method for analysis of soil organic matter using
and legions of biological interactions4). Fourier transform infrared absorption
The simplest way to evaluate organic matter spectrometry (FTIR). The spectra were
in soil is to measure total organic matter. Loss measured before and after pyrolysis, and the
on ignition, which is measured by combusting diŠerence between the two spectra, which
soil samples using an electric furnace, can be represented the organic matter in the soil, was
useful to estimate total organic matter, although used to compare soil samples. This method was
it includes a part of the crystalline OH of applied to four soil samples which were
minerals and strongly absorbed water. Wanogho indistinguishable by color and loss by ignition.
et al.28,29) proposed to use the weight loss from All four soil samples could be clearly
the decomposition of organic matter with diŠerentiated.
hydrogen peroxide, which is necessarily carried The usefulness of saccharide levels in soil
out prior to mineralogical examination. has been discussed for a simulated crime scene11),
Demmelmeyer60) proposed that total carbon but the method has not yet been thoroughly
content determined by using wet combustion studied for forensic science purposes.
with potassium bichromate would be
representative of total organic matter.
102 Yoshiteru Marumo

10. Biotic Materials plant cells for use in criminal investigations and
10.1 Plant Fragments pathology. They utilized characteristic features
Small fragments of plants in soil samples of epidermis cells of plants, especially those of
can be informative for forensic soil food plants, to identify the stomach contents of
identiˆcation. Plant fragments are also homicide victims71).
frequently found on clothes or car ‰oors, 10.2 Plant Opals
providing useful information about a link If soil samples are brought from diŠerent
between a suspect and a crime scene. If leaves are sites but have similar mineralogical
in a good condition of preservation, it is not compositions, they cannot be distinguished by
di‹cult to identify their species. The leaf petrological techniques. Plant opal examination
epidermis is composed of epidermis cells, can be a useful approach to solve such a
subsidiary cells, stomata, and hairs with a problem, because plant opals in soils are
variety of shapes and arrangements; thus, dependent principally on vegetation and are
microscopic observation allows us to compare independent of geological materials72). Surface
plant fragment samples or identify species by soil contains silica particles called plant opals or
referring to an atlas. opal phytoliths, which are formed through
Hiroe66) examined a great number of progressive siliciˆcation of cells in plant bodies.
monocotyledonous and dicotyledonous plants in As plant opals are casts of plant cells, they have
Japan. In this study, specimens were prepared by a variety of unique shapes, especially those
peeling oŠ the lower epidermis with sharp formed in the monocotyledonous grasses have
tweezers and observed under a compound species-speciˆc morphology. Kondo, and Sase
microscope at a magniˆcation of 100400. and Kondo7375) have proposed a classiˆcation
Generally, however, plant chips that are system for gramineous plant opals comprising
encountered in forensic science laboratories are seven classes: festucoid (boat-shaped); panicoid
withered or partially decayed so that it is di‹cult (dumbbell-shaped); chloridoid (saddle-shaped);
to peel oŠ the epidermis. Marumo et al.67) used a bambusoid (acutely angled outlines); fan-shaped
scanning electron microscope (SEM) to identify (fan-shaped or rectangular outline); elongate;
plant fragments directly. After plant fragment and point-shaped classes.
samples were softened by boiling in water, they Generally, light particles with a density of
were dipped in ethanol to remove the water and less than 2.3 separated by heavy liquid
stretched on a ˆlter paper for drying. Then, the (bromoform/ethanol) from the 0.010.2 mm
fragment was cut into two pieces, and one of fraction are subjected to observation for plant
them was turned upside down so that both sides opals under a compound microscope at
of the leaf could be seen on the same SEM magniˆcations of 100400. Sugita has simpliˆed
sample holder. the procedure to use the ＜0.05 mm fraction,
Westerkamp and Demmelmeyer68) from which clay minerals have been previously
published an atlas of foliar leaves from trees and removed by sedimentation, instead of using the
shrubs growing in Central Europe with 250 heavy-liquid separation technique. These
scanning electron micrographs and two procedures can also be used to prepare soil
diagrams. It includes a list of their samples for examination of diatoms.
morphological characteristics, species 10.3 Diatoms and Microfossils
distribution, and habitat requirements. An atlas Diatoms are frequently observed in soil,
of scanning electron micrographs of Japanese since they can rapidly propagate in various
Gramineae grass was published by Marumo et environments even during temporary wet
al.69) It includes pictures of both upper and lower conditions. Because of the distinctive diversity of
epidermis at a magniˆcation of 100500 of 89 diatom species and assemblages, diatoms can
species. Bock et al.70) published a guide to food provide useful information for forensic soil
 Forensic Examination of Soil Evidence 103

identiˆcation72). A series of articles about other hand, pollen assemblages from diŠerent
diatoms entitled ``Diatoms: diatom microscopy'' places, even with similar vegetation, showed
appeared in the journal Microscope76). signiˆcant diŠerences within the same
Regarding Japanese diatom ‰ora, a textbook geographic region. They also conˆrmed the
originally written by Kawamura and edited by validity of pollen analysis for comparison of soil
Ueno77) is useful. These resources will be helpful samples from shoes with those from places
to forensic scientists who are interested in where shoeprints were impressed90). Bruce and
diatoms as trace evidence. Dettmann91) discussed pollen/spore assemblages
Diatom tests have also been used for the in surface soils of Queensland, Australia.
postmortem diagnosis of drowning78). Although Several case reports describing the
there have been many reports on the use of successful application of pollen analysis to
diatoms to diagnose drowning7986), the details criminal investigations are available. Horrocks
will not be discussed here. and Walsh92) reported a rape case to which pollen
Soil where sand from alluvial or diluvial analysis was applied. Soil samples taken from
deposits has been used for construction may the jacket and pants of a suspect were compared
contain microfossils. Kurata and Hirano87) with soil at the crime scene and soil from where
found small cylindrical particles in soil samples the suspect's car was parked. The suspect had
when they were dealing with evidence from a insisted that although he certainly had been
criminal case. Detailed study revealed those there, he had stayed beside his car without
particles to be Bryozoa fossils, which had been in running after the victim. But the results of the
the sand materials used in the construction of a pollen analysis strongly supported the suspicion
building. Such unique fossils can be useful for of guilt because the pollen assemblages in the
determining the location where the soil evidence samples from the suspect correlated with those in
originated. the soil of the crime scene.
10.4 Pollen and Spores In the case of a private airplane crash near
Criminal investigations require forensic Ruidoso, New Mexico, in 1989, the problem was
pollen analysis to establish the presence or to determine whether a small mass of biotic
absence of localized pollen species in soil materials isolated from the fuel assembly had
evidence. For this purpose, species of forensic formed before or after the accident. In this case,
interest are generally insect pollinated, and not pollen analysis and soil examination played an
wind pollinated. Pollen of insect-pollinated important role by showing that the biotic
species tends to remain near the site of materials had formed during storage of the
deposition of their ‰owers, whereas wind- crashed airplane9398).
pollinated species disperse pollen long distances 10.5 Microbiology
and distribute it over an extensive area. A useful Although it has been said that soil is a
protocol was provided by Eyring88), who made treasury of microorganisms, reports about
some modiˆcations to traditional methods. microbiological techniques for forensic soil
Eyring describes a detailed procedure for pollen comparison are very rare. Thornton et al.99)
specimen preparation for microscopic proposed a biochemical approach to forensic soil
observation, and includes important references comparison. They proposed to measure the
about pollen extraction and identiˆcation, activity of phosphatase, arylsulfatase, urease,
pointing out that pollen in soil is sometimes trypsin, and invertase by colorimetric and
stained or has altered features. titrametric methods as essential indices of
Variation in pollen assemblages in surface biological activity in soils. The Michaelis
soils from site to site was examined by Horrocks constant (Km ) is considered to be particularly
et al.89). Samples from within the same localized signiˆcant because it is independent of sample
area showed a high degree of similarity. On the size. But it should be noted that because enzyme
104 Yoshiteru Marumo

activities ‰uctuate and shift with time, the soil re‰ected in the composition of those oxides,
evidence and exemplar soil must be collected at allowing us to diŠerentiate soil samples. The
approximately the same time. application of various instrumental analyses of
Omelyanyuk et al.100) proposed a elements in soil has been summarized by Bain et
multisubstrate testing method for forensic soil al.103).
comparison. In this method, 11 carbon sources Thermoluminescence (TL) is the emission of
such as lactose, acetate, etc. are used for light when a material is heated to temperatures
incubation of microorganisms in soil extract, just below incandescence. TL curves recorded as
and the growth of microorganisms is monitored light intensity versus temperature depend on the
by using the color reaction with tetrazolium particular materials involved and their minor
violet. components and on environmental history such
as radiation and thermal and pressure
11. Instrumental Analysis conditions. Ingham and Lawson104) studied the
Studies on the application of elemental application of TL to forensic soil comparison.
analysis to forensic soil identiˆcation have been Samples were exposed to radiation from cesium
attempted using xray ‰uorescence spectrometry 137 or cobalt60, and then TL curves were
(XFS). Hiraoka101) studied discrimination of measured. The results for 630 samples showed a
soils with energy dispersive XFS using 110 soil coincidental match of only 2.3.
samples collected from Kyoto prefecture in High performance liquid chromatography
Japan. Statistical evaluations revealed that (HPLC) of organic compounds in soil has been
concentrations of rubidium, strontium, studied by Nilsson105) and Andrasko106). They
potassium, and iron showed good correlation proposed that the analysis of polycyclic aromatic
with geological materials at the sampling sites. hydrocarbons (PAH) would be useful for
These results suggest that elemental analysis by forensic soil identiˆcation. PAHs, which have
XFS could be useful for determining the original two or more benzene rings, are produced by
location of soil evidence. combustion or pyrolysis of fossil fuels and other
As Thornton4) has pointed out, the organic materials. An analysis of PAHs in
interpretation of elemental data will remain systematically collected soil samples showed
somewhat clouded because of the heterogeneity signiˆcant discriminating power. Reuland and
of elements in soils. Elemental analysis requires Trinler107) studied HPLC analysis for forensic
relatively large amounts of sample (several soil identiˆcation and emphasized that if
grams) and is not applicable to trace soil constituents that exist only under certain
evidence. Since elements exist in diŠerent forms ecological conditions are found in soil samples,
in the soil, it is preferable to use an analytical that evidence will greatly assist in police
method speciˆc to those chemical forms. For investigations.
forensic science purposes, Marumo102) studied a Cengiz and Sakul108) reported on the
simpliˆed method that used XFS to determine application of anion analysis to forensic soil
free oxides of iron, aluminum, and silicon in discrimination. Capillary electrophoresis was
soils. A dithionite-citrate system buŠered with applied to soil extract. Chlorite, nitrate, sulfate,
sodium bicarbonate (Mehra and Jackson's and phosphate contents were compared among
method) and an acidic ammonium oxalate soils.
system (Tamm's method) were used to extract A successful pH measurement method for
diŠerent type of oxides. Mehra and Jackson's soil evidence using a colorimetric technique has
method can extract both amorphous and been developed by Dudley109).
crystallized oxides, and Tamm's solvent
dissolves amorphous oxides. The conditions
under which a soil developed are strongly
 Forensic Examination of Soil Evidence 105

12. Other dimensions.

12.1 Ceramics and Concrete Benko114) described polarized microscopic
Hashimoto et al.110) reported an appropriate observation and FTIR analysis of dust. Cooking
method for forensic identiˆcation of ceramics. grease, vegetable oils, some candle wax
Their results showed that sulfuric acid etching formulations, and certain semivolatiles may
yields favorable results for micromorphological provide deˆnitive spectra. Residues from
observation by SEM. cleaning/polishing products also gave unique IR
In a murder case, a corpse was embedded in spectra.
concrete, and a bucket to which concrete was
adhered was found in a suspect's home. Visual 13. Case Studies
examination of cross-sections of concrete On 16 March 1978, the prime minister of
fragments under a binocular microscope and Italy, Mr. Aldo Moro, was kidnapped on his
elemental analysis by XFS successfully identiˆed way to the Parliament House. In the morning of
the concrete fragments. 8 May 1978, Mr. Moro's body was found in the
12.2 Dust trunk of a car. The results of the forensic
Because dust is a complex mixture of investigation were disclosed after 20 years.
various distinctive types of particles and ˆbers, it Lombardi115) discussed the ˆndings of the
can hold rich information for criminal forensic investigation conducted on the evidence
investigations. If a particular combination of found on Mr. Moro's clothes and shoes (beach
some unique components in dust can be sand, bitumen, vegetable materials, and
established, it can be strong proof of contact polyester fragments), and in/on the car. The
with a particular place. Millette and Few111) study of the vegetable materials suggested that
described ˆve basic dust-sampling methods they had been picked up at a time close to the
suitable for obtaining samples for microscopic killing. Thermosetting polyester, of the type
analysis, with illustrations of samplers and a list used in boat manufacturing, was found under
of 30 useful references on dust sampling. The the fenders, in the tires, and inside the car, as
ˆve basic sampling methods are brushing, well as under Mr. Moro's shoes, supporting
scraping, lifting with adhesive tape, wiping, and proximity of a beach. Pollen analysis showed
vacuuming. One-liter ziploc plastic bags are part that the adhesion of volcanic soil to the car
of an eŠective technique proposed for collecting fenders antedated the adhesion of the sand.
settled dust for microscopy in almost any In November 1989, an aircraft ‰ew from
situation112). The bag is used as a collection San Diego, California, toward Ruidoso, New
glove, and then the dust is preserved in the bag Mexico. The aircraft crashed a few moments
until examination. before a scheduled landing near Ruidoso in
Millette and Few113) also investigated indoor Lincoln County, New Mexico. The contribution
carbon soot particles using transmission electron of soil evidence to the investigation of this case
microscopy. They were able to classify carbon was reported by Daugherty98). Because the plane
soot into four groups: (1) carbonized wood or had been exposed to intense ˆre after the crash,
cellulose materials ranging from micrometers to the change in soil color by heating was examined
millimeters in size and often retaining some of by using a redness rating test, which can
the features of the original materials; (2) hard, illustrate the relation between reddening and
shiny porous or hollow carbon spheres, typically heating of soil samples.
10100 mm in diameter; (3) spherical or near-
spherical particles with mean diameters of 14. Conclusion
around 20 to 30 nm adhering to each other to Because sand and clay are core components
form straight or branched chains; and (4) of soils, their mineralogical examination is
spheroidal carbon particles of colloidal certainly inevitable. More important for forensic
106 Yoshiteru Marumo

soil identiˆcation, however, are screening tests Verlag, Berlin (1986).

of soil samples. Soil color and particle size 5) Demmelmeyer, H.:
distribution tests are easy to perform and have Bodenkunde/Mikrobiologie, Kriminalistik
high discriminating power when used in Handbuch f äur Praxis und Wissenschaft,
combination. Especially, soil color, in which Vol. 2, pp. 135149, Boorberg (1994).
both soil development conditions and 6) Andrasko, J.: Soil. In Chemical
mineralogical composition are re‰ected, has an Criminalistics (Eds. Maehly, A. and
importance almost equivalent to that of soil Str äomberg, L.). pp. 171181, Springererlag,
mineralogy in forensic soil identiˆcation. Soil Berlin (1981).
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essential for forensic soil examination. However, science of forensic geology. Geotimes,
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