6(4):606-612 3 2 & 6 606 Forensic Science 9 2 2 fe eee ee Forensic Botany: Using Plant Evidence to Aid in Forensic Death Investigation Heather Miller Coyle’, Cheng-Lung Lee”’, Wen-Yu Lin‘, Henry C. Lee"’, Timothy M. Palmbach' ‘Forensic Science Program, University of New Haven, West Haven, Conn, USA; *Criminal Investigation Department Hsinchu Municipal Police Bureau, Hsinchu, Taiwan; "Nuclear Science Department National Tsing Hua University, Hsinchu, Taiwan; ‘Forensic Science Center, Taipel City Police Department, Taipei, Taiwan; and “Henry C. Lee Institute of Forensic Science, University of New Haven, West Haven, Conn, USA Abstract Forensic botany is still an underutilized resource in forensic casework, although it has been used on ‘occasion, Its an area of specialty science that could include traditional botanical classification of spe- cies, DNA, or materials evidence (race and transfer evidence, crime mapping or geosourcing, all de pendent on the specific case application under consideration. Critical tothe evaluation of plant ev dence is careful collection, documentation, and preservation for later scientific analysis. This article reviews proper procerlures and recent cases where botanical evidence played a ole in establishing e ‘ther manner or time of death, Plant evidence can be useful for determining fa death was due to an ace dent, suicide, or homicide, of what time of year burial may have taken place. In addition, plant ev dence can be used to determine ifa crime scene isa primary or secondary scene and to locate missing bodies. Ultimately the value of botanical evi- dence in a forensic investigation will depend on ‘what transpired or was overlooked at the crime scene. Scientists will not be able to use their plant analysis to resolve a critical issue ifthe investigator fr crime scene technician failed to start the pro- ‘cess. At the cote of this foundation is a require- ‘ment that investigators’ recognize the relevance of plant material, properly document where it was found, establish its relationship to the surround ings, and conduct suitable collection and preser vation techniques (1) (Fig. 1]. Seemingly one of the ‘easiest tasks at a crime scene, recognition of perti- rent evidence, is in practice one of the greatest challenges. The genesis of this problem lies i reality that what is relevant at today’s scene may or may not be relevant tomorrow. Thus, neither insti- wwwem).hr tutional knowledge nor checklists will provide a crime scene investigator with the requisite skills, ensuring that all key evidence is recognized. Moreover, whereas experience is of great value, it alone will not dictate the relevance of a particular item of evidence to that specific investigation. In the case of botanical evidence, this problem is ex asperated by the general lack of knowledge of this, subject matter. Plant Evidence Collection and Preservation Overcoming a potential recognition problem is best accomplished by making crime scene personnel aware of the possible value of plant evidence and sharing successful case exam- piles (2-6). However, several of these case stories ineum Croat Med J 2005;46(4):606-612 Figure 2.4 schematic diagram of the proper sequence for forensic evisence. First and foremost, evidence must be recognized at the erime scene. Secondly, but lees In portantly, evidence must be well documented, Finally, co Tection ana preseruation must be maintined so 98 to pre- serve the integrity ofthe evidence. which plant evidence was properly utilized are probably attributed to the general methodology of ‘crime scene processing, Investigators ate trying to establish linkages or associations between the vic- tim, suspect, scene, and individual items as wi any form of evidence (1) (Fig. 2) view | <———> [sane IRs cy — Figure 2, Botanical evidence may be used to link suspects, stins, and erme scenes using four-way linkage theory. Thus, investigators would know the value of locating plant material on a suspect or vic- tim, and then establishing the origin from which that plant came, On a macroscopic level, this may be a reasonable outcome. A large branch of clump ‘of leaves found within a victim’s palm will catch the curious eye of an investigator. In contrast; What about botanical evidence at the microscopic level, such as grains of pollen? Only if an investigator is aware of the potential existence of that evidence will any efforts be made to search for it. Finally, with limited or no training in botany, crime scene personnel will likely not know whether or not the plant material is indigenous to the area, relatively ‘common, or rare. Thus, guidelines should dictate that, when in doubt, plant.materials should be collected along with all necessary control samples. Crime scene documentation is necessary to preserve the character and manner in which evi= dence was located and collected from a scene, and in showing the relationship of that evidence to the overall scene and other evidence. Proper scene documentation involves multiple formats, such as. general note taking, photography, video taping, and sketches and diagrams. Each form of docu- ‘mentation is meant to complement the other. Fur- ther, documentation must be conducted through- ‘out the entire scene process, and capture, as much a possible, the events leading up to recognition, as well as post collection. Whereas documenta tion is important for any type of evidence; docu: ‘mentation of transient or transfer types of evidence 's very important since they may be subject to change over time, Most plant evidence, with the exception of pollen, may deteriorate, dry, mold, or be otherwise altered from the form in which itwas first discovered. For example, a piece of vegetative ‘material that was recovered from the scene, but neither documented nor properly preserved may degrade to the extent that examination and classt- fication is limited. However, if sufficient quality documentation occurred it may provide additional formation for a forensic botanist in classifying the plant species. Also, good documentation could be of assistance to a subsequent case recon- struction in regards to how that particular piece of plant material was transferred to the body of the victim. Was it clearly placed or created by direct, contact, or could it merely have blown onto or otherwise relocated onto the vietim in a manner that is irrelevant? Finally, thorough overall scene documentation may assist the botanist in deter- mining if recovered plant material is potentially in ddigenous to an atea, given the general topography and ecology. ‘Only after recognition and documenta- tion should an attempt be made to seize and pre- serve plant material. However, it may be impera~ tive to make the seizure as soon as possible to pro- tect the sample from further degradation or con- tamination, Proper packaging will vary depending fon the exact nature of the material, but in most ‘cases botany specimens should be packaged in pa- per after drying to prevent bacterial growth or ex- cessive moisture which would further degrade the ‘material. Separate items should be packaged sepa- rately to avoid, cross Contamination. The precise location where each item was located should also 607 ye 42 ao Auejog pur seuss aus,Coyle et al: Crime Scenes and Botany 608 Croat Med J 2005;46(4):606-612 be recorded. Finally, since this evidence is indeed evidence in every sense, it must be collected and maintained in a manner in which a chain of cus tody is established. Simply, the evidence must be accounted for from the moment it was first seized Until the completion of the case, and perhaps be- yond in the example of cold case re-evaluations. Also, the location and flow of evidence transfers must be logged, and these transfers made only when necessary. Another consideration is the need for control samples. Investigators must attempt to lo- cate any potential sources of the material at the scene or in adjacent areas, This can be a great chal lenge ifthe evidentiary pieces of plant material are so small that itis difficult for investigators to asso- ciate it with @ potential source based on visual characteristics. Further, if DNA typing isto be con- ducted on plant material, it will be necessary to obtain samples from numerous plants of the same species for the purposes of developing a meaning- ful data set for comparison and statistical analysis. All of this collection needs to be done contempo- aneous with the original scene or time of transfer as plants are subject to change over a relatively short period of time. Whereas their genetic pro- files will not vary over time, their physical atrib- tutes may vary greatly over different seasons. Case Examples Homicide or Suicide? Two cases in Tai- pei, Taiwan, show how plant material can be very simply used to establish whether an unidentified body was a result of homicide or suicide. In the first case, the body of a young woman was found lying in a gutter in an urban section of Taiwan. Be- fore autopsy, due to lack of obvious bone frac tures, itwas thought that the victim died as a result of ahit and run accident. She was visible on a sur veillance system tape, but after a truck passed by. her, she was no longer visible on the survetllance tape. It was assumed thal she was struck by the ve- hicle and her body moved to the gutter to hide the accident. Her body had already been relocated to the hospital by the time investigators reached the crime scene, so they decided to go to the hospital to examine the body. Some plant material (a tiny berry and stem) was found in the victim’s hair that ‘was unusual flora for the area, possibly from the genus Solanaceae, based on limited morpholog cal characteristics (Fig, 3B). After searching crime scene for any plant belonging to the genus Solanaceae, part of a broken stem was found at the location where the body had been discovered. In- vestigators looked upwards and found some pot- ted plants on the edge of a railing above the gutter (Fig. 3A), Those plants were identified as Solanum nigrum L., consistent with the botanical evidence from the victim's hair and there was a physical ‘match between the stem on one plant and the stem fragment found in the gutter (Fig. 3C). Based on lo- cation and climate, wind force would not be Figure 3. A. Photograph of the alley where the vetin’s body Was found ara the buildings from where the victim jumped. B. A tny berry and stem fragment found in We Young woman's hair C, Physical maten between the bro ken end of the stem fromthe alant on the rling anc the broken stem fom iNet eumeum Croat Med J strong enough to break the stem, so investigators presumed the plant had received a strong impact from something heavy such as a body. Also, the height of the railing where the plants were found ‘was 3.5 meters so they could not be reached by persons walking on the street. The likely scenario. for this incident was that the female fell from the top of the building, her body made contact with the plants during the fall, and some of the plant ‘was transferred to the gutter and the victim's hair Several days later, autopsy results showed her death to be a result of her impact injuries and her relatives told the police that the young woman sut- fered from depression and had attempted suicide before, Another case from Taipei, Taiwan, used similar botanical evidence to aid in the determina tion of manner of death. A male’s body was found hanging in a tree in a wooded area. It was clear that his death had resulted from asphyxiation due to hanging; however, the question was whether he had hung himself or if it was a result of foul play. The investigators at the scene noted that he had moss smeared on the insides of his wrists and when they examined the identical moss growing ‘on the tree’s branch, realized that he must have transferred the samples to his skin while tying the rope to an upper branch. His death was determi- ned to be a suicide (Fig. 4) Figure 4. The moss found on the vet’ wrists was eor= sistent with that on anatne location ofthe tree where the Tope was ted, indicating he had hime ted the rope and then died Primary and Secondary Crime Scenes. With any type of crime, plant evidence may be used to link a weapon or a body back to a primary crime scene, The following exemple shows how plant evidence can be used to identify the primary ‘or secondary scene. The bady of a young man was found in the guiter on the side ofthe road in Ta 2005;46(4):606-612 wan, with plant material grasped in his hand. Many injuries were evident including some contu- sions just below the knees. At autopsy, a fragment of bamboo was recovered from his stomach con- tents. In the gutter, where the body was found, no bamboo was growing, The bamboo evidence was used to locate the scene where the victim, injured, survived on water and bamboo before he crawled in the gutter for help, and eventually died (Fig. 5) The fact that he had grass in his closed hand indi- cated he was alive atthe time he was in the gutter. Later information revealed that he had been a vic~ tim of a hit and run accident and was being taken to the hospital when the guilly persons changed their mind and left him for dead in the bamboo ‘5, Abamboo laf that wae re2ov tins stomach contents after autopsy, indicating he spent ‘some timein a nearby bamboo grove before erawiingta the area where his body was ultimately discovered, Inan insurance fraud case, a young man, desperate for money, decided to use a cleaver to amputate his own hand and claim insurance money (Fig. 6). The insurance money would result from him claiming that he had been a victim of a robbery while driving his truck and the robbers. Figure 6. Grass recovered ‘rom inside, the newspaper lused to wrap the man’s hard led the investigators to a nearby area where jt besame evident trat he nad severed his own attempt to 609 P9160 Aareog pure sou auCoyle et al: Crime Scenes and Botany 610 Croat Med J 2005;46(4):606-612 had cut off his hand, His hope was that he would be compensated by the company for his perma rent disability. Although there was a lot of blood at the scene where he had pulled his vehicle over to the side of the road, investigators were suspi- cious due to the angle of the cut that severed his hand. After further investigation of the vehicle, they found some grass fragments wrapped within a bloody newspaper. They searched the surround ing area and found the location where his hand had actually been severed; a low stone wall in a nearby wooded area. Here, the same grass species ‘was growing as was found in the newspaper which he later confessed he had used to cover his arm ‘while getting back into his vehicle to call for medi- cal assistance, The grass samples linked the ampu- tation to a different scene and refuted his alibi Another case example used algae to link weapon used in a homicide to a nearby freshwa- ter pond, In the Hoepplinger case, a young wife ‘was murdered by her husband in the living oom of their home (7). Her head had been struck by a blunt object, and plant material and abrasions ‘were on her body and clothes. Initially, her hus- band insisted she was found on the sofa, but when the plant material matched to other vegetation in the driveway, he changed his story. Further inves- tigation led to the identification of a brick in the pond behind the house with hair and tissue from the victim. How could the husband be linked to the weapon and the scene? The answer was in the slain on the T-shirt that he had been wearing when he said he found his wife's body (Fig, 7A), Micro scopic analyses by botanists and forensic scientists identified the algal species on the shirt as being the same as those found in the pond, thereby estab- lishing the link between the husband and the ‘weapon (Fig. 78). Plant Succession and Clandestine Graves. Land that has been disturbed will follow a set pattern of plant re-colonization called plant succession. When an area-has been cleared of plants, such as-for'@ shallow grave site, then the primary colonizers are grass, followed by small shrubs and finally tees. Depending on the sun- lightshade and soil conditions, one can estimate the time of death at these sites based on the plant growth. A simple example of this is the postmor tem dating of a skeleton located in a field by the age of a plant found growing through the eye socket of the skull (4). This type of evidence can be Figure 7. Algae linking aweapon used in a homies toa nearby freshwater pond. A. Stained Tshirt the husband was wearing a the ime of the erime, nanging over the ral ing of hs home. B.A photomieragraph of the algae present fon the slained areas of the T-shirt that was tne same as found inthe pond. used to give a minimum estimate ofthe time; how- ever, not a maximum estimate, as there is no cer- tain manner to determine precisely when the plant seed was initiated relative to the skeletal remains. Inoother words, the skeleton could have lain in the field for some time prior to when the plant sprouted. Inaddition, scientists with experience in taxonomy and ecology can aid in missing persons cases. For example, a father and daughter were last seen together in Nevada, but a short while fater, the father was found dead in his vehicle due toa selfinflicted gunshot wound (7), The only evi- dence of the daughter was her jacket with some leaf litter covered in blood on the seat of the vehi- cle. The leaf litter represented greenleaf man- zanita, canyon live oak, interior live oak, ponder- cosa pine, black oak, and white fir, The growth re- quirements including Sunlight, knawn geographic distributions, water, and elevation were used to eumeum Croat Med J 2005;46(4):606-612 “profile” where this combination of tree and shrub species could be located, and five potential sites ‘were selected to search for the missing girl. At the fifth site, atan area that was a few miles past where the vehicle was found, her body was located, hidden between two logs Time of Death. Many different tests can be performed to establish time of death, including the use of body temperature, forensic entomology, and forensic botany. After death occurs, the body will slowly cool to the surrounding temperature. Before the body reaches the equivalent ambient temperature, itis fairly straightforward to calculate time since death by extrapolating back from the ‘original average body temperature. Both forensic botany and entomology may prove useful tools for estimating time of death. Entomology relies on predictable pattems of insect colonization of a corpse to estimate time of death (8). Forensic bot. any, however, can be used to establish time of death by pollen analysis (7,9) or analysis of stom ach contents (7). A good example of pollen analy- sis and time of death estimates comes from a study of skeletons recovered from a mass grave in Magdeburg, Germany, in 1994 (5,7). Since 32 male skeletons were recovered, it was hypothe. sized that they died in a military interaction. Two possible hypotheses were considered: (a) the men. ‘were victims of the Gestapo at the end of World War Il in the spring of 1945 or (b) they were Soviet soldiers killed by secret police after the German Democratic Republic revolt in summer of 1953. ‘The remains were exhumed and the nasal cavities rinsed with a saline solution to recover any pollen that the men may have breathed in prior to their death. Appropriate control samples from the soil were also analyzed for pollen species composi- tion. Seven of the tested skulls contained high amounts of plantain, rye, and lime tree pollen — all, ‘common for plants that flower in the summer _months, thus supporting the second hypothesis. The analysis of stomach contents can also be used to estimate time of death using simple microscopic methods (7). For example, a man in Colorado was suspected of being murdered. in 1993 by his wife. He was the eighth husband of il Caroll and his body was found in his home. The victim was very habitual in that he always ate the same breakfast consisting of toast, hash brown po- tatoes and eggs atthe samerestaurant every morn- ing. At autopsy, his stomach contents were mis identified as noodles, and then later correctly identified as potatoes. Knowing the estimated di- gestion time for this food, the victim must have died sometime between two and four hours after breakfast; a time frame that was not consistent with the wife's alibi, She was later convicted of the Summary ‘The recognition, collection, and preser- vation of botanical evidence are crucial to the suc~ cess of subsequent laboratory testing. Without the firs steps of the process being performed properly, the legal acceptance of the evidence into court ‘may be called into question. As with other forms of forensic evidence, plant material may be exam- ned using simple microscopy of more sophisti- cated plant DNA testing to enhance the informa tion gained from an investigation. Analogous to other forms of forensic testing, forensic botany first requires the identification of the plant species by morphological characteristics, microscopy, or mo- lecular biology (2-6). Alter the species has been identified by its class characteristics, then an at- tempt to individualize the sample is made. The ef- fectiveness of source attribution of plant material will depend on how unique the species is to the geographic area and its genetic history (7). If a plant is very rare, source attribution may not be difficult; however, many plants may require DNA. testing to confirm that the evidentiary sample orig inated from a source plant, if generated by seed, or a source population if generated by clonal repro- duction (7). Ultimately, the information from gen= erating plant DNA databases can be used to corre late specific markers with known plant varieties; to establish forensic linkages, to make patent appli- cations for new varieties; and to assess the degree of genelic-variaion found within a plant popula tion. This article discussed several cases where bo- tanical evidence was used to provide investigative leads or resolve cases and described the basic forensic principles of recognition, collection, and preservation of plants as evidence References 1 tee HC, Palmbach T, Miller MT, Henty Lee's crime scone handbook. San Diego (GA TAcaemie Pres 2 Bock!H, Nomis D6. Fatensiebotanyan underutilized resource. | Forense Se, 1997;42:3687 611 ye 42 ao Auejog pur seuss aus,612 Croat Med J 3 Hall DW. The contibutions ofthe forensic botanist 40 time seene investigations. The Prosecutor. 198822 4 Quattehomme G, Lacoste A, Ballet P, GevinG, Oller [A Contrbution of microscope plan anatomy to post ‘momar bone dating | Forensic Sc. 1997:42:140-. 5 Miler Coyle H, Ladd C, Palmbach T, Lee HC. The Green Revolation: botanical coniibuion to forensics and deug enforcement Croat Med J. 2001/42:3405 6 Wille P, Heilman A Estimating ime since death using plant roots and sms.) Forensic Se 198732126470, 7 Miller Coyle H,etor.Forensie botany principles and applications to criminal casework. Boca Raton (FL {Re Pres: 2005, 8 Gof i, Ay forthe prosecution: how insect evidence helps solve crimes. Cambridge IMA Harvard Univer sity Press; 2000. 2005;46(4):606-612 Sribor 8 Schubert C, Schoning R, Krause D, Wend U, Pollen analysis reveals murder season, Nature. 1998; Received: May 23, 2005 ‘Accepted: May 30, 2005 Correspondence to: Heather Miller Coyle Forensic Science Program University of New Haven 300 Boston Post Road ‘West Haven, Conn, USA plantdnatracker@yahoo.com