International Journal of Volume 1 Issue 2 Criminal Investigation 87-94

DEFINING THE ELEMENTS OF NEW SCIENTIFIC DISCIPLINES - PALYNOFORENSICS

Emilian M. DOBRESCU1*, Gabriel Ion OLTEANU2, Edith SIMA3

1) Romanian Academy, Bucharest, Romania 2)„Al. I. Cuza” Police Academy of Bucharest, 1A Aleea Privighetorilor, Bucharest-014013, Romania 3) “Spiru Haret” University Bucharest, Romania

Abstract Featuring the latest advances of science, forensics is constantly renewed by adopting the latest methods, procedures and techniques for identifying facts and everyday phenomena. The study of is one of these new methods, which greatly helps in forensics. Our paper deals with presenting the latest results in forensic palynology. We propose that palinoforensics be a new scientific discipline of the border, studying forensic evidence based on pollen.

Keywords: Forensic pollen; palynology; palynoforensics.

Introduction Pollen is a very fine powder form of Abundance - Pollen is produced in the anthers of flowers. It is composed of the anthers of flowers. Most flowers use grains of different shapes and colors wind or insects and small animals to help characteristic of each plant. It differs in transport their pollen to the female part of a form outer surface, the content of different flower on another plant of the same species, nutrients, vitamins, etc. Pollen can be so they must produce enormous amounts of obtained by direct harvesting by humans pollen (100-100,000 grains per anther) to from plants that have large amounts or by make sure some of it reaches its intended bees. Unlike pollen collected directly from destination. BUT – most pollen ends up as humans, using bee pollen is more valuable particulate components of soil, dirt, dust and because it is gathered from many flowers rocks. Therefore, pollen is everywhere. (Polyphemus) being added to bees, nectar Resistance to degradation - Pollen and saliva. Pollen carries the male sex cells and spore walls are so resistant to of flowering plants and plants that produce degradation that if they are deposited in the cones (eg pine trees). Spores are asexual right conditions they can be preserved in reproductive bodies of ferns, mosses and rocks for millions of years. Complexity – fungi. There are four major characteristics most plant species produce pollen or spores of pollen that make it a useful scientific that are different from pollen of other plant tool: species. This enables palynologists to link Microscopic size - most pollen grains dispersed pollen and spores found in rocks, are 10-70 um in diameter (there are 1,000 soil, dust etc., with the plants that produced um in a mm). them.

* Corresponding author: [email protected]

87 E. DOBRESCU et al.

Due to the development of properties over time. nanotechnology applications, plant pollen Pollen develops when a single can be used to identify perpetrators of chemically oxidized silicon in contact with crimes carried out with firearms. titanium oxide or mixed oxide. “Specific Microscopic pollen grains may remain composition of the mixture may vary attached to the cartridges, even well after depending on the weapon and the cartridges the bullet was fired and went through such “said Paul Sermon, professor of a barrel at high speed. criminology at the University of Surrey. Each “nanomark” is made up of According to UK Home Office in pollen grains and can be identified by Britain are held each year about 18,489 chemical composition analysis. Pollen crimes by armed attacks. Firearms and grains belonging to different species of ammunition are naturally contaminated with plants such as lilies, having a diameter of microscopic particles of pollen. Nanomarks about 30 micrometers are invisible to the can be used to solve crimes with an alternative human eye. Hundreds of pollen grains may method of harvesting the latest generation of remain attached to a cartridge, for periods skin cells from the surface of the cartridge. exceeding several years. The first such Cells remaining on the gun and cartridges nanomarks consist of ordinary pollen, a epidemic are the remains belonging to a dead natural substance that evolution has person who has handled the cartridge last. endowed with extraordinary adhesive

Forensic and palynology Palynology is the science that deals Artificial classification takes into with the study palynomorphs (included in account only ornamentation and categories of vegetables or having morphology of pollen and spores - Potonié vegetable affinity), as well as spores, type classification. Palynology has pollen or prepollen. Palynology branches applications in many areas: taxonomy, are: a) current palynology paleoclimatology, archeology, geology, (actuopalynology) or palyno-logy in situ stratigraphy, the study of honey study pollen and spores extracted from the (Melissopalinology) oenology (oenolo- structure of plant breeding and gical), especially in forensics (forensic paleopalynology, palyno-morphs studying science) Palynology shares common features fossils. with the paleoecology and ecology is Palynology in situ using a natural particularly useful in researching problems classification palinomorfs- spores and problems of these sciences. pollen bearing the name in accordance with Paleopalino-logia (Palynology) is the Code of Botanical Nomenclature. There closely related with Palaeobotanical (field are classifications of palynology - an of study of macroscopic plant remains high, artificial classification system which and paleoalgology (study of fossil algae) follows a natural parallel to the taxonomic shares also Paleopalynology study with hierarchy (parataxonomy), due to the various groups of algae. ignorance of the taxonomy of origin spores and pollen.

88 International Journal of Criminal Investigation, 1, 2, 87-94 DEFINING THE ELEMENTS OF NEW SCIENTIFIC DISCIPLINES - PALYNOFORENSICS

Legal Palynology dealing with the and has even been proposed as an additive study of pollen and dust, establish that a for bullets to enable tracking them. body or other object was in a certain place For instance, a dead body may be at a time. found in a wood, and the clothes may Pollen can tell a lot about where a contain pollen that was released after death person or object has been, because regions (the time of death can be determined by of the world, or even more particular , but in a place other locations such a certain set of bushes, will than where it was found. That indicates that have a distinctive collection of pollen the body was moved. species (Bryant V.M.). Pollen evidence can Today, New Zealand is the world also reveal the season in which a particular leader in the use of forensic palynology, and object picked up the pollen (Mildenhall the acceptance of this type of evidence in D.C.,). courts of law. Pollen has been used to trace activity In most of these cases the palynomorph at mass graves in Bosnia (Wood P.), catch data were an important factor in either solving a burglar who brushed against a Hypericum the case, or they were used to identify and link bush during a crime (Mildenhall D.C., 2006) a suspect to the scene of a crime.

History elements of forensic palynology use

Early researchers in palynology were The case in Sweden revolved around a the Greeks, and the first researchers’ woman who was killed in May, during a trip known names - Marcello Malpighi and in central Sweden. During the court hearing, Grew Nehemjah (in the seventeenth a number of experts, including a century). Palynology is a science in the palynologist, were asked to examine dirt 1836, through the work of Goeppert. In the attached to the woman's clothing. The twentieth century a well known name objective of those studies was to determine related to palynology is Ehrenberg. if the woman was killed at the same place Palynology is represented by the modern where she was found, or if she had been Couper, Faegri, Erdtman. In Romania, killed elsewhere and then dumped at the palynology was founded in Cluj by Emil scene where her body was discovered. Pop. Preliminary studies of the pollen in the It is difficult to establish precisely dirt samples suggested that she had been when the field of forensic palynology was killed elsewhere because the dirt lacked born. Attempts, whether successful or not, pollen from plants common in the area made prior to the 1950s, probably did not where the body was found (ie Plantago, gain much public attention and therefore Rumex and grasses). were not reported. Or, it is possible that if However, a later reinterpretation of the earlier attempts were made, the results may forensic pollen samples noted that the have been purposely hidden from the murder could have occurred in May because media in order not to alert criminals about that was before the grasses and herbs in the the use of this new technique. Two of the region had pollinated. The two opinions earliest reported cases using forensic were both used as evidence in the court palynology occurred in 1959 - one in Sweden proceedings, but we do not know if the and the other in Austria (Erdtman, 1969). murder was ever solved. http://www.ijci. eu eISSN: 2247-0271 89 E. DOBRESCU et al.

The importance of this case is that it In other early cases, during the 1960s is one of the earliest records in which and 1970s, Max Frei, a noted Swiss pollen data were considered as important criminalist, often used pollen as a forensic forensic evidence in a court case. tool to link suspects to events or to crime The second case, based primarily on the scenes (Palenik, 1982). Some of his most evidence recovered from a pollen sample noted cases include one in which a suspect associated with the crime, which occurred claimed that his pistol could not have been in Austria, dealt with the discovery of the used to commit a recent murder because it murdered victim's body, and the conviction had not been removed from its storage box of the criminal. During a vacation along the in months. However, Dr. Frei proved the Danube River, a man disappeared near suspect was lying because grease on the Vienna, but his body could not be found. pistol contained alder and birch pollen, both The police soon found a suspect having a of which were pollinating when the murder motive for killing the missing person, but occurred, not when the suspect claimed he had no evidence to link the person with the had last cleaned the pistol and put it away. possible crime. Without a confession or a In another case Dr. Frei showed that a body, the prosecutor's case seemed document was a forgery because he found hopeless. As the investigation proceeded, a fall-pollinating cedar pollen stuck to the ink search of the suspect's room revealed a pair used to sign a document, which had a June of boots with mud still attached to the date (Newman, 1984). Max Frei also gained soles. These were taken as evidence and fame for his pollen analysis of the Shroud of given for analysis to Wilhelm Klaus, a Turin, which revealed that the Shroud had geologist with the Austrian Geological probably been kept for some time in Israel Survey. Dr. Klaus examined the mud and and Anatolia (Wilson, 1978). found that it contained modern spruce, The term “forensic palynology” refers willow, and alder pollen. In addition there to the use of pollen and spore evidence in was a special type of 20 million-year-old, legal cases (Mildenhall, 1982). In its Miocene-age fossil hickory pollen grain broader application, the field of forensic present in the mud. palynology also includes legal information Based on the pollen evidence, Dr. Klaus derived from the analysis of a broad range was able to pinpoint where the defendant must of microscopic organisms--such as have walked while getting mud on his boots. dinoflagellates, acritarchs, and chitinozoans- Only one location, a small area 20 kilometers -that can be found in both fresh and marine north of Vienna along the Danube Valley, had environments (Faegri et al., 1989). soils that contained the precise mixture of However, in most sampling situations pollen found in the boots' mud. forensic palynologists rarely encounter When confronted with the identity of these other types of organisms because most this location, the shocked defendant are restricted to fossil deposits. confessed his crime and showed the More recently, the forensic analysis of authorities where he had killed the victim pollen grains has been used as a theme in a and then buried the body, both of which fictional murder mystery novel called occurred in the precise region pinpointed Probable Cause (Pearson, 1991). by Klaus.

90 International Journal of Criminal Investigation, 1, 2, 87-94 DEFINING THE ELEMENTS OF NEW SCIENTIFIC DISCIPLINES - PALYNOFORENSICS

According to the author, the In spite of all the television coverage identification of specific pollen grains and media attention, both the prosecution found, attached in the earwax of a and defense missed one potentially valuable murdered person was significant enough piece of evidence - forensic pollen evidence evidence to link the suspect to the murder, that might have been attached to the and to “swing the jury and win a defendant's clothing. Had the clothing that conviction.” During most of 1994 and Simpson supposedly wore on the night of 1995, one of the most publicized court the crime been examined, it might have cases in history made front-page headlines contained certain types of pollen that the in newspapers and magazines all over the prosecution could have used to link the world. This was the murder trial of the suspect to the scene of the crime. If the famous football and movie star OJ examination revealed no pollen, that piece Simpson that ended in October, 1995 when of evidence could have been used by the the jury found the defendant innocent. Mr. defense to argue that the defendant was not Simpson was accused of killing his wife and at the scene of the crime. Testimony during one of her friends during a rage of jealousy. the Simpson trial suggested that the person, News stories reported that the trial became as or persons, who committed the double popular as a media event that over one and a murder may have hidden in bushes in front half million people watched it daily on TV. of the Simpson home waiting for the During those court proceedings, some of the victims. If this assumption is correct, it is most important items of evidence were possible that pollen from the flowers on the forensic samples of fibers and hairs, and bushes, or pollen that may have fallen from DNA analyses of blood-stained clothing and the flowers onto the bushes' leaves might blood found at the , in the have brushed off on the assailant's clothing. defendant's car, and in his home.

Methodology palynoforensics Pollen and spore production and Many aquatic angiosperms live dispersion are important considerations in completely submerged and release their the study of forensic palynology. First, if pollen underwater, relying on water currents one knows what the expected production to transport the pollen from the male anther and dispersal patterns of spores and pollen to the female stigma of a neighboring (called the pollen rain) are for the plants in flower. This method of transport, like the a given region, then one will know what wind, is a hit- and -miss method of type of “pollen ” to expect in pollination. For this reason, these plants samples that come from that area (Bryant, produce pollen types that consist only of a 1989). Therefore, the first task of the single-layered cellulose wall, the pollen is forensic palynologist is to try to find a almost never preserved in sediments and match between the pollen in a known generally oxidizes rapidly if removed from geographical region with the pollen in a water. Because of these limitations, these forensic sample. Knowledge of pollen types of pollen are of little potential value dispersal and productivity often plays a for forensic work. major role in solving such problems. Another small group of plants are There are different methods by which called “autogamous” because they are self- plants disperse their pollen or spores. pollinating and are so efficient that little pollen is needed. http://www.ijci. eu eISSN: 2247-0271 91 E. DOBRESCU et al.

Pollen from these plants is rarely This last point covered proves both dispersed into the atmosphere even though advantages and disadvantages. Thus, on the their pollen preserves well and has a one hand if the pollen of a zoogamous plant durable outer wall, called an “exine,” made is found in a forensic sample, there is a of a stable chemical compound called high degree of reliability that the pollen “sporopollenin.” Like pollen produced by belongs to the forensic sample and it is not submerged plants, the pollen of an atmospheric contaminant. autogamous plants is of little value in On the other hand, so little pollen is forensic work because it is dispersed in produced by each plant that the chances of minimal numbers. its pollen getting into a forensic sample are In a larger group of plants, called reduced. The last category is the wind- zoogamous plants, pollination is dependent pollinated (anemophilous) type. This group upon the transport of pollen by some type includes a wide range of producers such as of insect (ie, bee, wasp, beetle, moth, ant) the gymnosperms and a significant number, or animal (ie, hummingbirds, lizards, but not the majority, of the angiosperms. nectar-feeding marsupials and bats, or This group also includes the spore- other small mammals). producing plants such as fungi, ferns, and Because of the efficiency, pollen mosses. productivity is low, yet not as low as if Because wind pollination is the most found in autogamous plants. The potential inefficient method of dispersion, value of zoogamous pollen in forensic anemophilous plants must produce vast work is excellent for two reasons. First, quantities of light-weight grains that will zoogamous pollen grains have some of the travel easily with air currents. Some species most durable exines. This means their of wind-pollinated plants, such as marijuana pollen will often remain preserved in (Cannabis), produce as many as 70,000 deposits for long periods of time which is pollen grains per anther (Faegri et al., generally less susceptible to destruction 1989). When large fields of these than pollen grains dispersed by other anemophilous plants grow together, their methods. Secondly, zoogamous pollen is flowers can produce millions of pollen produced in low amounts, thus it is not grains that are daily dispersed during the normally a potential contaminate found in flowering period. the pollen rain of an area.

In many cases, this abundance have come from almost anywhere as a result becomes a disadvantage because often of “random air dispersal” of that plant's marijuana pollen occurs in trace amounts pollen. on the shoes of people connected with the An example of this occurred during drug trade. Nevertheless, when such the summer of 1995 when European evidence is found, a palynologist cannot newspapers reported that “clouds” of state in court that “traces of Cannabis Cannabis pollen were drifting across the pollen could only have come from direct Mediterranean from source areas in association with, or use of, the actual Morocco, where local farmers reported plant”. Instead, if asked, a palynologist growing a bumper crop of marijuana. would have to admit that traces of European residents along the Mediterranean marijuana pollen on a suspect's shoes could coast were also warned by local newspaper

92 International Journal of Criminal Investigation, 1, 2, 87-94 DEFINING THE ELEMENTS OF NEW SCIENTIFIC DISCIPLINES - PALYNOFORENSICS

not to breath “too much” of the Cannabis free throughout the storage, laboratory pollen because it could cause extraction phase, and the analysis process. It hallucinations. This last statement, is also important to keep accurate records of however, is completely false because how each sample is collected and what has Cannabis pollen does not contain any of the happened to each sample from the time of hallucinogenic cannabis. collection until the completed analysis and Nevertheless, the high atmospheric report are presented. To ensure the court counts of Cannabis pollen illustrate how validity of forensic evidence, it is some of these grains might accidentally compulsory that a palynologist be able to occur in some forensic samples. state under oath that the materials, and the Another important factor is the subsequent pollen samples collected from “sinking speed” or rate at which a pollen those materials, were stored in a locked and grain falls to earth. Marijuana, alder, secure location. This aspect is also essential juniper and birch pollen are very small and in order to avoid the accusation that very light. Their average fall rate is about someone else, who was not authorized, had 2cm per second. On the other hand, maize access to a sample and “switched” it. If any plants and fir trees produce pollen that is kind of contamination or switching, either large and heavy, and falls to earth at a 15 natural or intentional, can be proven or times faster rate than the lighter ones. implied, then doubt will be cast upon the Using only these two examples, one resulting interpretations (Skinner et al., can notice that the potential distribution 1988). One final concern is the amount of area of maize and fir pollen grains will be material that will be collected for forensic smaller and more restricted that the analysis. In most cases very little dirt, mud dispersion area covered by the pollen from or other debris is available for collection plants in the first category (Tauber, 1967). and analysis. Therefore, most forensic In forensic studies this means that palynologists face several immediate when maize and similar types of large problems. Firstly, they will generally not pollen grains are found in samples, small have enough samples to experiment with dispersion areas are indicated and greater different collection techniques in order to precision in identifying the source region determine which one works best. Secondly, may be possible. The way in which they will often not have enough samples to samples are collected and processed is also conduct a second test if something goes critical, and both must be done correctly in wrong (ie, a centrifuge tube breaks, a beaker order to obtain accurate results. Ideally, spills, or a microscope slide breaks). pollen samples should be collected by a Thirdly, sample size may be further reduced competent palynologist who is when other potentially useful tests may knowledgeable in forensics. Such experts need to be carried out first (ie, soil will know how to collect contamination- testing, searches for fibers, sand grain free samples, and will know what analysis) before a destructive pollen precautions should be taken in order to analysis can be attempted. ensure that samples remain contamination-

http://www.ijci. eu eISSN: 2247-0271 93 E. DOBRESCU et al.

Conclusions

A 2009 report conducted by the Agricultural plant pollination by British Ecological Society and presented at insects leads to a cumulative annual market a workshop in the auditorium of the of vegetable food worth more than $ 22 University of Leeds, warns an ultimatum billion. Every year, India produces around calling on the serious dangers which would 7.5 million tons of vegetable foods, which arise if populations continued to decline in propelled it as a second place in the world pollinating insects galloping the pace in the after China. However, this huge amount can last two decades . not cover basic food needs of the Indian The worst situation is that the decline population. World Food and Agriculture in pollinating insects directly produces a Organization (FAO), affiliated to the UN, dramatic reduction in agricultural estimates that of the approximately 100 production, leading, ultimately, to reduce species of vegetables, grains and legumes, agricultural production from those plants which provide over 90 percent of food for and then to famine. Researchers have used 146 countries on the planet, about 70 that argument and study reports submitted species are pollinated by wild bees and by their colleagues in India, according to other remaining insect species. which, in recent decades, the insects that In this context we suggest that pollinate cereals, vegetables and legumes palynoforensics be considered a new of vital interest to the public, have begun to scientific border discipline, studying disappear. forensic evidence based on pollen.

References

Bryant V.M., Forensic Palynology: A New Walsh K.A.J., Horrocks M., Palynology: Its Way to Catch Crooks, position in the field of , (http://www.crimeandclues.com/pollen. Journal of Forensic Sciences, 53, 5, htm) January 22, 2011 2008, 1053-1060. Mildenhall D.C., Wiltshire P.E.J., Bryant Wiltshire P.E.J., Forensic Ecology, Botany, VM, Forensic palynology: Why do it and Palynology: Some Aspects of Their and how it works, Forensic Science Role in Criminal Investigation, Criminal International, 163, 3, 2006, 163-172. and Environmental Soil Forensics, 2009, Mildenhall, D. C., Hypericum pollen 129-149. determines the presence of burglars at Wood P. Pollen helps war crime forensics, the scene of a crime: An example of (http://news.bbc.co.uk/1/hi/sci/tech/364 forensic palynology, Forensic Science 0788.stm), January 22, 2011. International, 163, 3, 2006, 231-235. Stackhouse R., Forensic studies look to pollen,(http://www.thebatt.com/2.8526/for ensics-studies-look-to-pollen-1.1224833), June 10, 2011.

94 International Journal of Criminal Investigation, 1, 2, 87-94