A TALE OF TWO CORCHORUS SPECIES: JUTE AND ITS SUBSTITUTES IN COMMERCIAL GOODS By Barbara Leahy Fallon A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of Forensic Science Master of Science 2016 ABSTRACT A TALE OF TWO CORCHORUS SPECIES: JUTE AND ITS SUBSTITUTES IN COMMERCIAL GOODS By Barbara Leahy Fallon Natural fibers from j ute ( Corchorus capsularis and C. olit orius ) are common in commodities such as cordage, sacking, and textiles. C urrently neither chemical nor microscopic methods exist to differentiate fibers or the single - celled fiber ultimates from the se two jute species. Observation of physical and optical properties in this work, including a comparison of the measured length, width, and area of ultimates, revealed no differences between the species. A novel , rapid, and simplified investigation of cellulose orientation - specific Raman ba nds was also insuffici ent to identify potential differences in the angle of cellulose microfibril orientation in the cell walls. Jute can be identified and distinguished from other common vegetable fiber on the basis of its microscopic characteristics. Furthermore, other natu ral fibers and some synthetic fibers may be substituted in commercial goods advertised as jute . Twenty of 113 (18%) of commercially obtained jute goods analyzed herein contained non - jute fibers, the most frequent of which was polyester. Non - jute fibers ap peared in textiles, sacking, and flooring products but not in geotextiles, cordage, or miscellaneous products advertised to contain jute. Photomicrographs of the commercial goods will be assembled into a pu blic ly accessible database. A summary of products containing non - jute fibers from the database may also help guide fiber examiners toward a more thorough examination of the jute product types known to more frequently contain non - jute fibers, resulting in a more efficient use of time and resources. Further , this visual database can help standardize interpretation of vegetable fiber features among fiber analysts. iii Dedicated with great love to my parents, Patricia Leahy Fallon and Peter Barry Fallon, for their selfless sacrifices, love, and encouragement. iv ACKNOWLEDGEMENTS school forever. It seems like yesterday that I first arrived in Michigan, excited, nervous, and determined. While my original plan for my time in Michigan underwent a few revisions, I am happier and stronger for having made this path and story on my own. M y committee has provided the tools to accomplish this project. I am grateful for ber. His - Counterfeiting and Product Protection summit this year afforded me an informative glimpse into a world few forensic scientists ever see. Second, I thank my advisor, Professor Ruth Smith, for more than three years of encouragement. Her trust and confidence i n me allowed me to pursue new subject matter for the MSU Forensic Chemistry Lab. I n doing so I was able to solidify my decision to pursue a career as a mic roscopist and trace evidence analyst . At every turn Ruth has been generous with her time, level - headed when I needed an anchor, and down - to - earth as a mentor. Many successes have happened when the right person appeared at the right time and was willing to take a chance on someone. I am exceptionally grateful for my final committee member, Dr. Chris Palenik, for being one of those people for me. His willingness to bring me into the Microtrace family as an intern resulted in several highly educational months with some indebted. At Microtrace, I profusely thank Skip and Chris Palenik for sharing their expertise, patience, and lab space with me. erving vegetable fibers, this project v would never have been conceived. Ethan Groves provided invaluable discussion s about vegetable fibers and le d the Raman data acquisition. Lab mates Katie White , Brendan Nytes , Ethan Groves , Katelyn Hargrave , and Jay Bec kert made me feel welcome and were also generous in sharing their time and knowledge with me, both in and out of the lab. I am also in awe of the incredible scientists at The McCrone Research Institute in Chicago who spent two weeks training me in PLM, hai memorabilia meant I was able to sit in the desk and read the notes of É mile Chamot, my academic great - grandfather, a truly inspiring experience . At Michigan State, I thank Amy Porter at the MSU Histopathology lab, who kindly provided free paraffin embedding and cross sections of reference jute samples free of charge. Professor Per Askeland provided initial consultations for Raman work. Professor A r deshir Azadnia graciously allowed me to borrow h all that well for glycerin jelly. Ruth approved my request to bring two undergraduates into the lab, Brittany Butterworth and Joshua Champine. Their patience as I learned how to teach made for a pleasan t first real experience with mentoring . Brittany prepared many slides for the commercial goods. Josh assisted with the macroscopic and microscopic photography of commercial goods , some slide preparation, and too many miscellaneous tasks. I further earnestl y thank my lab mates, past and present, to include Jordyn, Alex, Trevor, Kristen, Fanny, and A few other individuals and organizations have made this research project possible. The MSU Pharm/Tox Department and my other thesis advisor, Professor Michelle Mazei - Robison, many outstanding individuals of LARA and the Quantico La b helped me keep thing in vi perspective as I worked toward my career goals. I also thank Jeff Dake, I think, for being another mentor who put a little faith in me and then promptly saddled me with terrifying but very exciting responsibilities. Lastly, I mus t acknowledge the organizations which sponsored this research. Microtrace provided reference materials, instrument time, general supplies , and work space for much of this project. The generous Cartha (Deke) DeLoach fellowship from the J. Edgar Hoover Found ation and the Society of the Former Special Agents of the FBI permitted me to live in Illinois during my months with Microtrace and purchase many reference books, commercial jute goods, and other supplies for this project. The MSU Graduate School provided Research Enhancement and Travel Funding awards for my attendance at SAFS/ASTEE 2015 and MAFS 2015, respectively. The MSU Council of Graduate Students also provided a Conference Grant to attend MAFS 2015. The MSU Forensic Science Program provided funding fo r supplies in the summer of 2015. I am a firm believer that nobody gets anywhere in life alone. Without my dear friend for Laura Fox, my best friend and wifey from Cornell University, occasionally checking in to screw my head back on straight, I could not have accomplished as much as I have, in school and to succeed, and egan. late mom, who taught me everything I know about being conscientious, candid, and compassionate: your water girl finally did it. Now onward, to that thing ca lled a career! vii TABLE OF CONTENTS LIST OF TABLES i x LIST OF FIGURES x KEY TO ABBREVIATIONS x i i CHAPTER 1: Introduction 1.1 The need for vegetable fiber identification 1.2 The botanical origins of jute fibers 1.3 Obtaining fiber from jute plants 1.4 The jute economy 1.5 Analysis of fibers in the modern crime laboratory 1.6 Microscopy of vegetable fibers 1.7 Raman microscopy of vegetable fibers 1.8 Aims of the present study 1 1 2 4 6 7 8 11 13 CHAPTER 2: Materials and Methods 2.1 Reference samples 2.2 Commercial samples 2.3 Sample preparation 2.4 Reference slide preparation 2.5 Microscopic observations 2.6 Measurement of ultimates 2.7 Qualitative test for degree of lignification 2. 8 Macroscopic photography 2.9 Photomicrography 2.10 Raman spectroscopy of cell wall cross sections 14 14 14 1 5 15 15 16 16 16 17 18 CHAPTER 3: Results and Discussion Part 1: Microscopic and spectroscopic attempts to differentiate ultimates of C. capsularis and C. olitorius 3.1 General morphology of C. capsularis and C. olitorius ultimates 3.2 Measurement of C. capsularis and C. olitorius ultimate dimensions 3.3 Optical properties of C. capsularis and C. olitorius ultimates 3.4 The Herzog test for the cellulose twist direction of C. capsularis and C. olitorius ultimates 3.5 Raman spectroscopy of C. capsularis and C. olitorius ultimate cell walls 3.6 New considerations for the Herzog test 19 19 21 23 28 31 35 CHAPTER 4 : Results and Discussion Part 2 : Microscopic analysis of commercial jute goods 4.1 Rationale for the analysis of commercial goods analysis 4.2 Acquisition and summary of commercial goods 3 8 38 38 viii 4.3 Features used to identify jute and non - jute fibers 4.4 Analysis of textile samples 4.5 Analysis of geotextile samples 4.6 Analysis o f cordage samples 4.7 Analysis of flooring samples 4.8 Analysis of sacking material samples 4.9 Analysis of miscellaneous samples 4.10 Trends in jute fiber substitution in commercial goods 4.11 Difficulties encountered in the analysis of commercial go ods 4.12 Photography of commercial goods samples for the future development of a jute fiber database 39 44 48 49 49 49 52 52 54 5 6 CHAPTER 5: Conclusions and Future Work 5.1 Efforts to identify a method to discriminate jute fibers from C. capsularis and C. olitorius 5.2 Evaluating the frequency with which goods advertised to contain jute are composed of other fibers 58 58 59 APPENDI X 62 BIBLIOGRAPHY 9 4 ix LIST OF TABLES Table 1 . Comparison of ultimate dimensions reported in this work and in a representative sample of published literature . 22 Table 2 . Summary of commercial goods analysis by product type . 39 Table 3 . Summary of the results of the phloroglucinol test for relative degree of lignification for reference samples depicted in Figure 14 . 41 Table A.1 . Description of items in the commercial goods collection and results of microscopic analysis . 63 Table A . 2 . Observations of fibers from the commercial goods collection . 84 x LIST OF FIGURES Figure 1 . A multi - scale representation of jute fibers . 3 Figure 2 . Enlarged photomicrograph of a representative ultimate of flax ( Linum usitatissimum , Sample B - 30008), a common bast fiber, to demonstrate the appearance of nodes, dislocations, and cross markings . 10 Figure 3 . Representative ultimates from reference samples of C. olitorius (A, Sample B - 30059) and C. capsularis (B, Sample B - 30025) . 20 Figure 4 . Measurement of ultimates from C. capsularis and C. olitorius reference samples . 21 Figure 5 . Optical properties of a representative ultimate from a reference sample of C. capsularis (Sample B - 30026) . 24 Figure 6 . Optical properties of a representative ultimate from a reference sample of C. olitorius (Sample B - 30007) . 25 Figure 7 . Enlarged photomicrograph of representative ultimates from C. olitorius (Sample B - 30007) to demonstrate the appearance of nodes, dislocations, and cross markings . 26 Figure 8 . Enlarged photomicrograph of crystals in an ultimate from a jute coffee bean sack (Sample BF - 009) . 27 Figure 9 . Photomicrographs of a representative ultimate of C. capsularis (Sample B - 30026) to demonstrate the Herzog test for a Z twist fiber . 29 Figure 10 . Schematic illustration of how the Herzog test for a Z twist fiber can result in either a definitive or uncertain determination of the twist direction . 30 Figure 11 . Structure of cellulose . 33 Figure 12 . Average Raman spectra of C. capsularis and C. olitorius . 34 Figure 13 . A new hypothesis to explain why some ultimates produce clear colors and others produce ambiguous colors during the Herzog test . 36 Figure 14 . Photograph of the results of the phloroglucinol test for relative degree of lignification for reference samples of jute, some jute s ubstitutes, and some common vegetable fibers . 40 xi Figure 15 . Ultimates from a sample of jute flooring (Sample BF - 012) to demonstrate the variability in lumen morphology observed in many of the commercial goods . 4 4 Figure 16 . Blend of polyester fibers from a swatch of fuchsia textile advertised as - 023) . 45 Figure 17 . Photomicrography series of a polyester fiber from a textile swatch - 024) . 46 Figure 18 . Blend of macerated fibers from coffee bean sacking (Sample BF - 005) and comparison to macerated coir fibers (Sample B - 30002) . 51 xii KEY TO ABBREVIATIONS DSLR Digital single - lens reflect GC Gas chromatography IJSG International Jute Study Group IR Infrared MSU Michigan State University n Refractive index n Refractive index parallel to the length of the fiber n Refractive index perpendicular to the length of the fiber NE - SW Northeast - southwest NW - SE Northwest - southeast PLM Polarized light microscopy PPL Plane polarized light SEM Scanning electron microscopy SWGMAT Scientific Working Group for Materials Analysis TLC Thin - layer chromatography XP Crossed polars 1 CHAPTER 1: Introduction 1.1 The need for vegetable fiber identification Forensic fiber examinations frequently attempt to associate a questioned fiber with a known source ( 1 , 2 ) ; this involves identifying the fibers present in each sample. Aside from noting the general prevalence of fibrous materials in the everyday environment, very few published works comment on the frequency with which fiber evidence is encountered in casework ( 1 , 2 ) . One trace evidence analyst noted that of 506 cases he examined between 1977 and 1983, 100 cases, or nearly 20%, yielded fiber evidence of investigative value ( 3 ) . While often overlooked and not collected at crime scenes, such trace evidence can be of particular value when other types of evidence have been exhausted or are absent ( 1 , 4 - 8 ) . Thus, accurate identification of fibers in casework can be exploited to provide helpful investigative leads. Jute is the most abundant vegetable fiber in production after cotton ( 9 , 10 ) and is used to make the well - known textile burlap . Currently neither micro scopic nor chemical methods exist to differentiate fibers from the two jute species, Corchorus capsularis and C. olitorius . Distinguishing between jute species would provide a new level of discrimination when jute fibers are encountered in forensic casewor k. Furthermore, other natural fibers and some synthetic fibers are sometimes substituted in commercial goods advertised as jute. In labs with limited time and resources, knowing which types of products are more likely to be misidentified can guide examiner s towards more thorough examination of particular product types. The experiments described herein represent one of the first attempts to discriminate fibers from the jute species and also determine the frequency of mislabeling of common commercial jute goo ds. 2 1.2 The b otanical origins of jute fibers Jute is the name for the fiber extracted from the bast (stems) of the jute plants, C . capsularis and C. olitorius . While approximately 80 species of plants belong to the genus Corchorus ( 11 ) , only these two are commercially cultivated for fiber production ( 9 , 12 - 17 ) . Jute fibers from C. capsularis can range from pale cream to dull gray or brown to black in color and be finer than those from C. olitorius , which is often yellowish, silky, and softer than the former type, although the fiber color is substantially impacted by processing conditions ( 12 , 13 ) . Jute plants are annual herbaceous angiosperms ( 10 , 13 ) belonging to the family Malvaceae ( 11 ) . Other fiber crops in this family include the seed fibers cotton ( Gossypium sp. ) and kapok ( Ceiba sp. ) as well as other bast fibers such as kenaf ( Hibiscus cannabinus ), roselle ( H. sabdariffa ), and aramina ( Urena lobata ) ( 11 , 18 ) . The high content of lignin in jute cell walls causes the fiber to be relatively coarser than other, softer bast fibers such as flax ( Linum usitatissimum ), hemp ( Cannabis sativa ), and ramie ( Boehmeria nivea ) ( 19 - 22 ) . In the jute plant, fiber bundles arise from the actively dividing procambium and cambium in the stem ( Figure s 1A and 1B ) ( 23 ) . These rings of tissue develop into the vascular tissues that serve as a transport system for the plant ( 24 ) . The vascular tissues include water - carrying xylem and nutrient - carrying phloem ( 25 , 26 ) . The tissue that is harvested for commercial fiber products begins as phloem - associ ated fibers that eventually differentiate into sclerenchymatic support tissue ( 10 ) . This occurs in part via the continuous deposition of lignin into the secondary cell wall ( 24 ) . Lignins, a complex and diverse family of biopolymers, impart mechanical strength to the plant stem by thickening the cell walls of the mature sclerenchyma fibers ( 19 , 24 , 26 ) . Nearby parenchyma cells, a thin - walled cell type that adds bulk to the stem, may infrequently remain attached to the final fiber product and be detected microscopically from scrapings ( 27 ) . 3 Figure 1 . A multi - scale representation of jute fibers. A . Sketch of a transverse cross section of C. olitorius stem. Fibers are located in bundles primarily surrounded by phloem tissue. Modified from Catling and Grayson ( 27 ) . B . Plane polarized light (PPL) photomicrograph of a transverse cross s ection of C. olitorius embedded in paraffin wax. Sample B - 30059 from the Microtrace Vegetable Fiber Reference Collection ( 18 ) . 400X total magnification. C . A longitudinal schematic representation of the cell wall layer structure typically found in bast and wood fiber cells. The striations indicate the typical directio n of cellulose spiraling in each layer. The microfibril angle is the acute angle between the long axis of the fiber and the angle of cellulose in the S2 secondary cell wall layer. Modified from Ye et al . ( 28 ) . Epidermis Cortex Phloem Cambial zone Fibers Xylem 100 m S1 S2 S3 Primary A. B. C. 4 The cell walls of most angiosperms are broadly divided into two categories: primary and secondary cell wall s. The former is present in all cell types and begins to form during plant cell division, and it is continuously synthesized and reformed as cells expand. In contrast, secondary cell walls are only laid down in sclerenchyma after the cell reaches its final size ( 29 ) . Lignin is deposited on a scaffold of cellulose microfibrils ( 30 ) . The secondary c ell wall is divided into three layers based on cellulose orientation: S1, S2, and S3, with the outermost S1 adjacent to the primary cell wall and the innermost S3 adjacent to the lumen ( Figure 1C ) ( 29 , 31 ) . S1 and S3 are relatively thin with cellulose microfibrils laid down perpendicu lar to the long axis of the fiber ( 29 , 31 , 32 ) . By comparison, the S2 layer is considerabl y thicker and the cellulose microfibrils spiral at a steep angle to the fiber axis ( 29 , 31 - 37 ) . This angle varies in fibers from different plant species in both degree of steepness and in S versus Z direction ( 31 , 38 - 40 ) . Multiple techniques can measure this spiraling angle, including two dimensional X - ray diffraction ( 38 - 40 ) , scanning electron microscopy (SEM) of the exposed cell wall ( 32 ) , ellipsometry ( 28 ) , and Raman spectroscopy ( 3 3 - 35 ) . As the first two techniques were not used in this research, the y will not be discussed further. Raman spectroscopy will be addressed in Section 1.7 . 1.3 Obtaining fiber from jute plants The conditions in which jute plants are grown can impact the final quality of the resulting fiber. Jute is hand sown in loamy soil during the rainy season between February and May in India and Bangladesh ( 10 , 12 ) . Soil characteristics affect the fiber quality: sandy soils result in coarser fiber s while clay soils result in shorter fibers that do not separate from the stem easily ( 12 ) . Fiber quality also deteriorates if jute is grown repeatedly on the same land without crop rotation. S pacing between plants proves to be another consideration : plants too close will have 5 thin stems and poor fiber quality, while plants far apart will branch and a lso diminish quality. Thus, the crop must be weeded and thinned two to three times by hand during the growing cycle to ensure strength of the final product ( 12 ) . The ideal time to harvest the fiber crop is considered to be when the fruits ripen, often between early Jun e and late October ( 12 ) . Harvesting too early reduces the yield of fiber, while harvesting too late makes fiber s overly coarse and difficult to separate from the stem ( 12 , 13 ) . Traditionally all plants have been har vested by hand using sickles to cut close to the ground or by pulling directly from water - logged areas ( 12 , 13 ) . The stems can then be defoliated, which increases the speed and uniformity of the next process, retting ( 13 ) . Retting frees the fibers from the stem by soaking tied bundles of stems in slow - moving water. Bacteria and fungi decompose pectins and hemicelluloses that attach the fibers to the rest of the stem ( 13 ) . The flowing of the wate r removes dark - colored and acidic degradation products, resulting in higher grade fibers that are stronger and more lightly colored ( 13 ) . In contrast, stagnant water permits deterioration of fiber color and luster due to the buildup of fermentation products ( 15 ) . C. capsularis ret s faster than C. olitorius because it supports a larger population of aerobic and anaerobic microorganisms, but the presence of a periderm layer on the exterior of the stem in C. capsularis decreases the uniformity of retting ( 13 ) . As soon as possible after retting, the fiber must be physically separated from the woody stem remnants. Stripping is also done by hand. In this process, the butt end of the fibers is gently beaten with a wooden paddle or mallet to loosen the fiber from the stems. Grasping and shaking the butt ends removes the bulk of the woody stem parts, while jerking the fibers through the water several times removes the final upper stem portions from the fiber. After a sufficient period of drying, the fibers are transpor ted to a baling center. Here, fibers are sorted, given a 6 preliminary grade, and marked with the species and area of origin before being purchased for further processing or export ( 13 ) . 1.4 The jute economy The Inte rnational Jute Study Group (IJSG ) is an intergovernmental body comprised of India, Bangladesh, and the European Union . The group was established by the United Nations Conference on Trade and Development and seeks to promote the worldwide jute industry. IJS G data show that between 1994 and 2010, the only period for which centralized data are available, worldwide jute production has rise n and fallen cyclically. Output fell to a minimum of 1.9 million kilograms in the 1995 1996 season and peaked at a maximum of 3.1 million kilograms in the 1997 1998 season ( 41 ) . Mo st recently, in the 2009 2010 season, India and Bangladesh respectively produced 5 7.4 % and 41.6 remaining 1% of jute ( 41 ) . Second only to cotton in production ( 9 , 10 ) , jute has been used for a wide variety of goods. A non - exhaustive list of jute products includes cordage, sacking, textiles, draperies, upholstery, rugs, carpet backing, wall coverings, handicrafts, linoleum backing, soft luggage, mail bag s, webbing, geotextiles/soil savers, agricultural mulching, tarpaulins, padding cloths, roofing materials, electrical insulation, backing for automotive carpeting, automotive seat backing, automotive composite materials, biocomposites, and fire - retardant g arments ( 9 , 10 , 14 , 15 , 42 ) . The production of jute goods b y India and Bangladesh quintupled between 1991 and 2013, from 521.9M kilograms to 2,569M kilograms ( 41 ) . In that time, the use of jute for sacking increased from 38.9% to 58.1% of all manufactured goods, while carpet backing declined from 11.6% to 0.5% as it has been largely replaced by synthetic materials. The significance of jute 7 evidence from carpet backing may therefore increase because it has become rarer in recent years. As th e uses of jute in different goods shifts with time, such information may assist a forensic examiner in narrowing the time window from which a product is likely to have originated. 1.5 A nalysis of fibers in the modern crime laboratory Currently, the Organ ization of Scientific Area Committees Materials Subcommittee, overseen by the National Institute of Standards and Technology, is developing standards and guidelines for the forensic examination of fibers ( 43 ) . While these recommendations for procedures and reporting are in progress, the retiring Scientific Working Group for Mate rials Forensic Fiber Examination Guidelines is still, for the time being, the widely accepted standard for fiber examination ( 44 ) . Most recently updated in 2014, the Guidelines were originally published in 1999. The Forensic Fiber Examination Guidelines is divided into chapters on microscopy, visible and infrared (IR) spectroscopy, thin - layer chromatography (TLC) of textile dyes, pyrolys is gas chromatography (GC), and analysis of bulk fabrics and cordage. The Guidelines recommend combining methods which provide complementary types of information. To perform the examination most efficiently, it is recommend that an analyst commence with te chniques which provide the most exclusionary information. As all samples handled in this research were intact specimens of different fibers or products, there was no need to determine if there was an association of one sample to any other. Techniques whic h could assist with such associations, but not necessarily in the definitive identification of the vegetable fiber itself, include the analysis of fiber color using visible spectroscopy or TLC of dyes , and analysis of bulk items for physical matches or sha red class 8 characteristics such as warp and weft count. Of the remaining techniques, IR spectroscopy would be more appropriate for the identification of a man - made fiber, as the primary constituent of all vegetable fibers is cellulose. Next, pyrolysis GC wo uld be most helpful for polymer fibers that do not char at high temperatures, as vegetable fibers would. With the various drawbacks of these techniques, microscopical analysis is the ideal choice for the identification of vegetable fibers. 1.6 Microscopy of vegetable fibers Much has been written on the subject of using microscopes to identify the fiber type(s) present in a sample. Some of the earli est materials are reference books containing black and white photomicrographs with varying amounts of informa tion on the chemical and physical characteristics of ultimates ( 16 , 45 - 47 ) . Various editions of The Particle Atlas have enumerated unique characteris tics of the most common vegetable fibers along with color photomicrographs, with Volume V of the second edition containing a dichotomous guide to identification ( 22 , 48 , 49 ) . Other reference texts and educational textbooks also suggest schemes for fiber identification using a variety of methods ( 20 , 21 , 50 - 52 ) . The SWGMAT Forensic Fiber Examination Guidelines also describes an ana lytical scheme using microscopy, detailed below, primarily for the purpose of excluding or including a known fiber sample as a potential source of another sample ( 44 ) . The initial classification of a sample as a vegetable fiber, as opposed to a man - made, animal, or mineral fiber, may be made quickly based on observations while handling the sample, such as the texture, sheen, odor, and uniformity of the sample ( 20 , 44 ) . However, it is preferable to support such initial observations with low - magnification examination with a stereomicroscope. Overall construction of the sample should be recorded, along with phy sical 9 characteristics such as fiber length, relative diameter, luster, apparent cross section, damage, and adhering debris ( 44 ) . If the fibers under examination appear si milar after the preliminary examination, a comparison microscope may be used to observe physical characteristics such as diameter, color, presence of delusterant particles, surface characteristics, and apparent cross sectional shape ( 44 ) . Cross sections may be taken of fiber bundles and viewed with a light microscope to note their shape ( 27 , 44 ) . While the above applies to any fiber, vegetable fibers should additionally be searched for additional adhering plant tissues and c ells, presence of crystals, and relative degree of lignification , nodes, dislocations, and cross markings ( 20 , 44 ) . While some authors do not distinguish between nodes and dislocations or sometimes describe only one or the other ( 17 , 27 , 45 , 46 ) , they are in fact separate features ( 53 ) . Nodes are regions where ultimates appear to swell and often have a bend, akin to a knuckle or e lbow joint ( Figure 2 ) ( 53 ) . Dislocations occur transverse to the length of the fiber and usually appear as bright, birefringent discontinuities in the shaft ( Figure 2 ) . The origin of dislocations is unclear and has been hypothesized to be due to tension or slippage resulting from compression; this disagreement by botanists on the subject has been succinctly summarized by Catling and Grayson ( 27 ) . Dislocations occur more frequently in bast than leaf fibers and are absent from seed fibers ( 27 , 53 , 54 ) . Another feature of vegetable fibers is cross - markings, which are thought to be the remains or imp ressions of adjacent cell walls no longer attached to the fibers ( 27 ) . These simple lines crisscrossing the width of the fiber ( Figure 2 ) ( 27 , 45 , 46 ) . Most vegetable fibers at crime scenes are encountered as technical fibers, i.e. bundles of individual fiber ultimate cells that may also contain additional tissue from the plant ( 20 , 27 ) . In t he laboratory, a maceration process is used to digest the lignin holding ultimates together, 10 Figure 2. Enlarged photomicrograph of a representative ultimate of flax ( Linum usitatissimum , Sample B - 30008), a common bast fiber, to demonstrate the appearance of nodes, dislocations , and cross markings. A. The ultimate is first oriented to the vertical position of minimum brightness under XP. B. T he first order red compensator may then be added to the light path to observe these features. Dislocations occur frequently; cross marking are fainter than dislocations. Letters indicate the following features: N = nodes, D = dislocations, and C = cross markings. Ultimates were mounted in glycerin jelly and photographed at 100X total magnification. C N, D N, D C D N, D N, D D N, D C D C A. B. 11 releasing them so th at the single cells may be examined ( 20 ) . For crime scenes where only one or a few technical fibers are recovered, th e analyst can infer that characteristics of ultimates a re representative of the entire sample. Maceration permits measurement or estimation of ultimate dimensions and obse rvations of dislocations, cross markings, and relative lume n diameter to overall diameter ( 20 , 44 , 47 ) . Next, a determination of optical characteristics is carried out using polarized light microscopy (PLM). For vegetable fibers, the most important properties include refractive index dete rmination (either definitive or relative to the chosen mounting medium), sign of elongation, bi refringence, retardation colors, and the Herzog test for cellulose twist direction (S or Z). 1.7 Raman microscopy of vegetable fibers As mentioned briefly in Section 1.6 , the Herzog test is used to determine the direction of ( 20 , 36 ) . It is further possible to determine the angle of the cellulose twisting using a variety of methods ( 28 , 32 , 38 - 40 ) , including Raman spectroscopy ( 33 - 35 ) . This technique is based on single - wavelength excitation of ground state molecul es in which inelastic scattering of photons from the sample induces vibrational transitions ( 33 ) . Lignocellulosic samples have classically suffered from laser - induced fluorescence which swamps the Raman scattering si gnal ( 55 - 58 ) , although this limitation has been mitigated by the use of near - IR lasers, Raman - inactive mounting surfaces, and interferometry ( 55 - 57 ) . Despite this challenge , Raman microscopy provides useful information about the orientation of biological macromolecules, such as cellulose and lignin, which is not available through IR spectroscopy or other microscopic observation s ( 55 , 56 ) . S pecifically, Raman permits such examinations of individual cell wall layers from a 12 simple cross section, whereas SEM would require potentially damaging electron beam exposure or isolation of the layer of interest ( 32 , 33 ) . The use of Raman microscopy for cellulose orientation determination is based on the prese nce of bands in the spectr um that are sensitive to cellulose orientation ( 33 - 35 ) . In one study by Gierlinger et al . ( 34 ) , Raman spectra were collected from the tangential surface of wood fibers rotated from - 90° to 90° relative to the long axis of the fiber. Relative heights of peaks whose intensities v aried most as orientation changed were used to develop a quadratic regression to describe the relationship between the peak height ratios and orientation angle. Multivariate analysis using partial least squares regression then identified the wavenumber reg ions of the overall spectrum that most contributed to the orientation - related variance. These regions were then used to develop a model which could predict the cellulose angle from the Raman spectrum of fiber cross sections. The validity of this model for predicting microfibril angle from cross sections of fiber samples was then tested and confirmed by comparing predicted angles with those experimentally determined by X - ray diffraction. While this method involved more statistical analysis than is typically performed in routine forensic fiber examinations, the most significant advantage of Raman spectroscopic determination of cellulose angle compared to historic two - dimensional X - ray diffraction is the reduction in sample preparation time and the simplicity o f data acquisition. This achievement notwithstanding, an even more simplified procedure could aid forensic fiber examiners in using cell wall cellulose orientation to discriminate fibers from different species, a property that is not currently measured in forensic settings. 13 1.8 Aims of the present study The accurate identification of vegetable fibers encountered in forensic casework can be useful to determine whether commercial goods contain the fibers indicated on their labels. Such information may be particularly useful in cases of suspected counterfeiting. While most vegetable fibers originate from only one plant species, jute is unique in that it is harvested from two species. Thus, the experiments describe herein aimed to devise a rapid, simple, and inexpensive method to discriminate between fibers from the two jute species with the hopes that such a method could then be used to determine whether commercial goods contained one or both jute species. Subsequently , the incidence of non - jute fibers in go ods purporting to contain jute was determined regardless of the jute species present in an item. Aim 1: Attempt to use microscopic and spectroscopic methods to distinguish jute fibers from C. capsularis and C. olitorius . Aim 2: Ascertain the frequency with which commercial goods labeled as jute contain non - jute fibers. 14 CHAPTER 2: Materials and Methods 2.1 Reference samples Reference samples and macroscopic photographs of jute and other vegetable fibers were a generous gift from the Microtrace, LCC Vegetable Fiber Reference Collection ( 18 ) . 2.2 Commercial samples Good s were obtained from a variety of sources, as summarized in Table A.1 in Appendix A . Briefly, samples were obtained from online craft product and home goods retailers, big box stores in Michigan (Meijer), professors from Michigan State University (MSU) , Ebay, a Michigan coffee shop, a Michigan dollar store, and an online clothing a nd acces sories retailer. Products were purchased or obtained by donation from brick - and - mortar stores in greater East Lansing, Michigan geographical region. To remain within a modest budget, only one or two stores of a particular type (e.g. home improvement stores ) were visited and multiples samples of the same product were not purchased. Textiles, defined as woven cloth for craft, clothing, or interior design purposes, were the largest group of samples (n = 85 ). Other samples included geotextiles for agricultural and conservation purposes (n = 7 ), cor dage (n = 7 ), flooring (n = 6), sacking (n = 6 ), and novelty/miscellaneous items (n = 2). 15 2.3 Sample preparation or pulled technical fibers were collected from bulk reference and commercial sa mples for maceration to release fiber ultimates. Standard maceration conditions were utilized ( 20 ) . Briefly, samples were boiled in a bath of equal pa rts glacial acetic acid and 30% hydrogen peroxide for 4 - 5 hours. After the maceration, the fluid was poured off and samples were rinsed twice with distilled water and twice with 70% ethanol. Macerated samples were stored in sealed, labeled glass vials in 7 0% ethanol. 2.4 Reference slide preparation References slides were prepared using tungsten needles to spread ultimates across a slide with the aid of a stereomicroscope (Nikon SMZ645). Glycerin jelly (5 g gelatin, 30 ml deionized water, 35 ml glycerol, 0 .5 ml phenol) was prepared and used as the mounting medium ( n = 1.43). A cover slip was pressed down evenly over the sample, which was then allowed to cool and solidify. Three coats of nail polish were used to ring the edges of the cover slip to prevent dr ying and cracking of the mounting medium. 2.5 Microscopic observations Samples were observed with a polarized light microscope ( Olympus BH - 2 or Olympus BX - 51) under plane polarized light (PPL), between crossed polars (XP), and between XP with a 530 nm first order red compensator inserted into the light path . Observed physical and optical characteristics included overall morphology, surface texture, relative lumen diameter to overall diameter, interference colors, sign of elongation, direction of twist, and presence of nodes , dislocations, and cross markings. For synthetic fibers, refractive indices parallel and 16 perpendicular to the long axis of the fiber (n and n , respectively) were determined using the standard Becke line method using reference Cargil le oils of known refractive indices. Birefringence was calculated as n - n . 2.6 Measurement of ultimates Length and area were measured with ImageJ (National Institutes of Health) from PPL photomicrographs by manually selecting the shortest path through the ultimate and around the perimeter, respectively. Average width was calculated by dividing area by length. Averages of the three measurements for each species were compared using two - tailed, u - tests performed with Microsoft Excel 201 0 . 2.7 Qualitative test for degree of lignification Phloroglucinol reagent was prepared by dissolving solid phloroglucinol in the minimum amount of 190 proof ethanol necessary and adding an equal volume of concentrated hydrochloric acid. Cuttings of selected reference fibers and all commercial goods were pl aced individually in watch glasses. Upon the addition of phloroglucinol reagent, highly lignified fibers turn a deep magenta, partially lignified fibers turn lighter pink, and celullosic fibers do not exhibit a color change ( 20 , 21 , 50 ) . 2.8 Macroscopic photography Macroscopic photographs of commercial samples were captured using an EOS Rebel T3i 18.0 megapix el digital single - lens reflect (DSLR) camera with an 18 - 55 mm lens (Canon). 17 2.9 Photomicrography Photomicrographs for the commercial goods database were collected with the same EOS Rebel T3i 18.0 megapixel DSLR camera connected to the Olympus BX - 51 microscope using a 1.38X widefield T - mount adapter (Martin Microscope). Unless otherwise noted, samples are mounted in glycerin jelly ( n = 1.43). Photomicrographs were collected with fibers oriented in multiple positions with the 10X objective for 140X total magnification. The EOS utility - density 25 filter was utilized for all 160X photomicrographs to achieve optimal illumination for pho tomicrography. To improve the white balance of brightfield photomicrograph, a daylight blue filter was Photomicrographs of samples observed under XP u tilized an exposure compensation set to - 3 2 / 3 . Photomicrographs of samples observed under XP with the first order red compensator inserted used an exposure compensation of - 1. PPL photomicrographs were collected with ultimates oriented in the northeast - southwest (NE - SW) position of maximum brightness as determined by examination under XP . The XP and XP plus compensator photomicrographs were collected while the fiber was subsequentl y rotated into the northwest - southeast (NW - SE) position, vertical extinction, and horizontal extinction. As vegetable fibers only to go pa rtial extinction, these positions w ere determined to be the positions of minimum brightness and were not always orient ed at precisely 0 ° or 90 ° to the x - axis. 18 2. 10 Raman spectroscopy of cell wall cross sections Technical fibers of samples B - 30058 ( C. capsularis ) and B - 30059 ( C. olitorius ) were mounted in paraffin blocks. Thin (10 m) sections were sliced with a microt ome and mounted on glass slides. A tungsten needle was used to gentle remove an embedded fiber cross section from the paraffin for mounting on an aluminum slide. Spectra were collected from the 10 m cross sections mounted on aluminum slides with a n inVia Raman microscope (Renishaw) with a charge coupled device detector using the following conditions: Objective: 100X ( Numerical aperture = 0.9) Spot size: 1.064 m Excitation wavelength: 785 nm Exposure time: 1/10 s Grating: 1200 lines/mm Accumulations: 2 For each species, one spectrum was collected from four randomly chosen locations in the interior of the cell wall from a single cell for one reference sample of each species (B - 30058 for C. capsularis and B - 30059 for C. olitorius ). Repli cate spectra were individually baseline corrected (cub ic spline interpolation method), zeroed, and averaged to generate a representative spectrum for the cell wall of that species. 19 CHAPTER 3: Results and Discussion Part 1: Microsc opic and spectro scopic attempts to differentiate ultimates of C. capsularis and C. olitorius 3.1. General morphology of C. capsularis and C. olitorius ultimates The reference collection contained five samples of C. capsularis (B - 30006, B - 30024, B - 20035, B - 30026, and B - 3 0058) and four of C. olitorius (B - 30007, B - 30027, B - 30028, B - 30059) as well as an additional five samples from the genus Corchorus of undetermined species (B - 30103, B - 30120, B - 30220, B - 30221, and B - 30222). Because the focus of the first part of the research effort focused on identifying potential differences between the two species, only the samples of known species are described in the following observations . Much literature on vegetable fibers treats jute from C. capsularis and C. olitorius as interchangeable ( 10 , 17 , 21 , 45 , 46 , 50 , 59 , 60 ) , and it is true that they share many similarities. As with all bast fibers, the overall morphology of ultimate ce lls was not uniform but in general was characterized by a larger diameter in the middle part of the fiber which eventually tapered to both ends ( Figure 3 ). One of the most characteristic features of C. capsularis and C. olitorius ultimates is the variability in the diameter of the lumen relative to the diameter of the overall cell, as illustrated in Figure 3 . Within a single ultimate, a lumen may vary from occupying greater than 50% of the total diameter to being pinched so narrow ly as to appear nearly closed. However, not every ultimate demonstrates this characteristic. Ultimates in every macerated sample have a lumen of more uniform diameter that occupies roughly one - fifth to one - third of the overall diameter ( Figure 3 ) . This is in agreement with ob servations published elsewhere ( 22 , 27 ) . In some ultimates of both species with particularly wide lumens, leng th - wise striations similar in appearance to the texture of wood grain could sometimes be observed (data not shown). Thus, gross morphology does not differ between C. capsularis and C. olitorius . 20 Figure 3. Representative ultimates from reference samples of C. olitorius (A, Sample B - 30059) and C. capsularis (B, Sample B - 30025). Overall, fibers were widest along the shaft and tapered toward the ends, unless one or both ends were blunt due to breakage . These ultimates demonstrate d the variability in lumen diameter characteristic of jute ultimates. Thin arrows point to sections of narrow, pinched lumen while bold arr ows point to sections of wider lumen. Dashed arrows point to ultimates with more uniform lumen diameter. Ultimates were mounted in water and photographed under plane polarized light (PPL) at 40X total magnification. 100 um A. 100 um B. 21 3.2 Measurement of C. capsularis and C. olitorius ultimate dimensions Next, the length, apparent area, and average width of ultimates were measured from photomicrographs by calibrating ImageJ software to convert the number of image pixels into a physical distance or area. This was done to test whether such a rapid, simple, a nd inexpensive method could discriminate ultimates from the two jute species. Ultimates from each species did not significantly differ in length, apparent area, or width. ( Figure 4 , unpaired , two - tailed Figure 4 . Measurement of ultimates from C. capsu laris and C. olitorius reference samples. The ultimates from each species did not differ significantly in length ( A ), apparent area ( B ), or average width ( C ) ( - tailed t - test s). Length was measured from photomicrographs using ImageJ to manually click the shortest path from one end of an ultimate to the other end. Apparent area was similarly measured by manually clicking around the perimeter of the ultimate. Average width was c alculated by dividing the apparent area by the average width. Values are presented as mean ± standard deviation. B . C . A. 22 - tests , p > 0.05 ) C. capsularis measured 2,089 in length, 24,908 ± 11,492 m 2 in apparent area, and 12.0 ± 3.0 m in average width (mean ± standard deviation, n = 35). C. olitorius measured 2,008 ± 588 m in length, 24,888 ± 8,954 m 2 in apparent area, and 12.3 ± 1.9 m in average width ( n = 34 ). Most prior reports of ultimate size do not distinguish betwee n samples of each species ( Table 1 ) . Fo r example, Catling and Greyson tabulate fiber ultimate measurements from four other authors , none of which treat the species as distinct ( 27 ) . The range of lengths extends from 800 to 4,100 m, while Catling and Grayson report 600 to 5,300 m (average 2,170 m) for C. capsularis and 600 to 5, 300 m (average 2,040 m) for C. olitorius ( 27 ) . Table 1 compares the values found in this study with length and width measurements reported by other authors. The measurements reported herein for each species fall within the range of previously reported values Table 1. Comparison of ultimate dimensions reported in this work and in a representative sample of published literature. Author, Year Re ference Species Length, average (mm) Length, range (mm) Diameter, average ( m) Diameter, range ( m) This work - C. capsularis C. olitorius 2.09 2.01 0.9 4.8 0.6 4.4 12.0 12.3 8.0 18.5 8.2 18.0 Catling and Grayson, 2004 C. capsularis C. olitorius 2.17 2.04 0.6 5.3 0.6 5.3 18 20 9.6 26.6 9.3 - 32.6 The Textile Institute, 1975 C. capsularis C. olitorius 1.9 2.4 2.3 3.2 - 16.6 20.7 15.9 18.8 - Franck, 2005 Not specified 2.5 0.75 6 18 5 25 Isenberg, 1958 Not specified 2.0 1.5 5.0 - 20 - 25 Luniak, 1953 Not specified - 0.8 8 - 5 32 Montgomery, 1954 Not specified 2.03 1.5 5.1 23 20 - 25 von Bergen, 1942 Not specified 2.4 - 10 - 23 for jute. The large overlap in measurements for C. capsularis and C. ol itorius seen both here and by Catling and Grayson indicates that measuring the dimensions of ultimates is not a reliable method for discriminating fibers of the two jute species. 3.3 Optical properties of C. capsularis and C. olitorius ultimates When viewed in different positions between crossed polars (XP) and between XP with t he first order red compensator , it is possible to observe the optical properties of the jute ultimates ( 61 ) . The interference colors observed in samples between XP arise because light passing through the fiber vibrates at different speeds and wavelengths in two directions , parallel or perpendicular to the length of the fiber ( 61 ) , due to the regular arrangement of cellulose chains in the cell walls ( 53 ) . The arithmetic differen ce in the wavelengths, known as the retardation measured in nm, corresponds to a color on the Michel - L é vy chart of interference colors, where lower orders of colors correspond to samples with lower birefringence ( 62 ) . If the thickness of the fiber is known, the wavelength of observed interference colors can be used to calculate a numeric value for birefringence ( 62 ) . However, because jute cell cross sections are irregular polygons ( 27 , 45 ) , it is not possible to obtain an accurate value for the thickness at any given point on the fiber. Thus interference colors are described here only to give a qualitative idea of a All jute samples observed from bot h species demonstrated interference colors ranging from first order pale yellows or grays to second order vibrant blues, as seen in the representative photomicrographs in Figure s 5 B and 6 B . Further, all samples exhibited addition colors in the northeast - so uthwest (NE - SW) direction and subtraction colors in the northwest - southeast (NW - SE) direction indicat ing a positive sign of elongation (Figures 5C , 5D , 6C , and 24 6D ) . Thus, these characteristics again do not permit differentiation of fibers from the t wo jute species. Further examination of the ultimates between XP and with the compensator helps to enhance certain features which can also be observed in PPL, including nodes, dislocations, and cross - markings ( Figure 7 ). In general, the jute reference samples contained these features as reported previously ( 17 , 21 , 27 , 50 ) . However, some author s have noted that neither nodes nor Figure 5. Optical properties of a representative ultimate from a reference sample of C. capsularis (Sample B - 30026). A. Reference photomicrograph in PPL focused on the center ultimate pointing in the NE - SW direction. B. Ultimate in the center demonstrates pale yellow interference colors between XP. C. In the NE - SW position the ultimate exhibi ts addition of wavelengths into the second order bright blues and green - yellows between XP with the compensator in place. D. In the NW - SE position the ultimate exhibits subtraction of wavelengths to first order light - medium yellows. Together C and D illust rate that fibers of C. capsularis have a positive sign of elongation. Ultimates were mounted in glycerin jelly and photographed in PPL at 100X total magnification. A. C. B. D. 25 cross - markings are common in jute fibers ( 17 , 21 , 27 , 45 , 46 ) . O bservations made in this work are not consistent with such reports ; nodes were seen in all fibers and were common. In addition, cross - markin gs were noted to occur occasionally to very frequently. At present it is unclear if these discrepancies represent a difference in judgement between the authors or a true difference in the features of the fibers. S ome of the same jute specimens examined by Catling and Grayson were examined as reference samples for this work, demonstrating that subjectivity in Figure 6. Optical properties of representative ultimates from a reference sample of C. olitorius (Sample B - 30007). A. Reference photomicrograph in PPL focused on the center ultimate in the NE - SW position. B. Ultimate in the center demonstrates pale yellow interference colors between XP. C. In the NE - SW position the ultimate exhibits addition of wavelengths into the second order bright blues and green - yellows between XP with the compensator in place. D. In the NW - SE position the ultimate exhibits subtraction of wavelengths to first order light - medium yellows. Together C and D illustrate that fibers of C. capsularis have a p ositive sign of elongation. Ultimates were mounted in glycerin jelly and photographed in PPL at 100X total magnification. A. C. B. D. 26 Figure 7. Enlarged photomicrograph of representative ultimates from C. olitorius (Sample B - 30007) to demonstrate the appearance of nodes, dislocations, and cross markings . The ultimate in focus in the center of the image is the same as in Figure 4 but has been rotated to a position of extinction to highlight nodes, dislocations, and cross mark ings. Letters indicate the following features: N = nodes, D = dislocations, and C = cross markings. Ultimates were mounted in glycerin jelly and photographed under PPL at 100X total magnification. N, D N C C C N D D 27 interpreting features can make comparing observations of different authors difficult. Alternatively , since cross - markings are thought to be the remains or impressions from where walls of adjacent cells attached to the fiber cells ( 27 ) , perhaps differing methods of extracting fiber from the plant stems resulted in more or less frequent cross markings. One feature of jute ultimates observed in this study but not discussed in any of the rev iewed literature was the presence of non - birefringent crystal - like inclusions ( Figure 8 ) . These formations occurred in all samples of jute but were present in less than half the ultimates of any given sample . They seemed to occur inside the ultimates themselves as single crystals and chai ns of such ( Figure 8 ) . While they bear some resemblance to many - pointed cluster crystals , t hey might be more accurately described as clustered irregular polygons . However, they appeared slightly smaller than crystals reported by Catling and Grayson ( 27 ) and were not birefringent. Further, Catling and Grayson state that clus ter crystals occur in jute far more rarely than cubic and rhomboid crystals, although these latter crystals were not observed in intact or macerated Figure 8. Enlarged photomicrograph of crystals in an ultimate from a jute coffee bean sack (Sample BF - 009). Single crystals appeared in the middle section while a short chain was found to the left side of the photomicrograph. Notice also the abundance of dislocations and cross markings. The macerated sample was mounted in glycerin jelly and photographed under XP at 400X total magnification. A. C. B. D. 28 specimens ( 27 ) . It is possible that different crystal morphologies may only become apparent upon ashing a sample ( 27 ) . However, this procedure, which heats the carbon - based vegetable fiber until it turns to ash and leaves behind crystals, was not undertaken i n this work for two reasons. First, many forensic laboratories attempt to analyze and identify fibers without destructive methods. Although the type of crystals present can aid in identification ( 27 , 53 ) , this method is indeed destructive and may consume more sample than can be spared for the analysis. Second, the laboratory in which these inclusions were f irst noticed did not contain the apparatus to perform such ashing. While larger samples were available for this study, whether or not these inclusions were indeed the cluster crystals described in Catling and Grayson ( 27 ) was not determined. 3.4 The Herzog test for the cellulose twist direction of C. capsularis and C. olitorius ultimates As discussed in Section 1.2 , vegetable fibers have a thick S2 secondary cell wall made of cellulose that spirals at an angle to the fiber axis ( 29 , 31 - 40 ) . The Herzog test is a quick and reliable method to determine whether the cellulose spirals in an S or Z direction and is the same for all ultimates of a species ( 20 , 53 , 63 , 64 ) . The test involves aligning the fiber under cross polars to the two perpendicular positions at which it appears to be nearest extinction, inserting the compensator, and observing whether the fiber appears blue or orange in the perpendicular and vertical o rientations. The blue color at the vertical extinction position and orange color at the horizontal extinction indicate the Z orientation while the opposite pattern of colors (blue when horizontal and orange when vertical) would indicate an S orientation. C onsistent with other reports ( 16 , 21 , 50 ) , all jute fibers examined were determined by the Herzog test to have a Z twist orientation of cellulose cell wall spiraling ( Figure 9 ) . 29 Figure 9. Photomicrographs of a representative ultimate of C. capsularis (Sample B - 30026) to demonstrate the Herzog test for a Z twist fiber . A , C. The ultimate was oriented at the positions of minimal brightness under XP in the vertical and horizontal positions, respectively. B. When the compensator is inserted, the fiber appears blue in the v ertical position. D. The fiber appears orange in the vertical position. The ultimate here is the same ultimate depicted in Figure 5 . The macerated sample was mounted in glycerin jelly and photographed at 100X total magnification. A. C. B. D. 30 During the course of observing many reference jute specimens, it became apparent that sometimes the blue and orange colors we re very easy to distinguish while in other cases the colors were ambiguous ( Figure 10 ) . For example, first the jute ultimate would be oriented into the horizontal extinction position. Upon inserting the first order red compensator, some fibers Figure 10. Schematic illustration of how the Herzog test for a Z twist fiber can result in either a definitive or uncertain determination of the twist direction. First, the ultimate is oriented to the horizontal position of minimum brightness under XP (top). When the first or der red compensator is inserted into the light path (bottom), a Z twist fiber is expected to appear orange. In some instances, this was clearly observed (bottom left). Other ultimates appeared mostly magenta with a very faint orange hue that could turn to magenta with a faint very blue upon the slightly rotation of the fiber (bottom right). The circle represents the field of view. 31 would clearl y appear orange while others appeared mostly magenta with only a faint hint of orange, which could change to the opposite color , blue , upon the slightest rotation of the fiber. It is important to remember that vegetable fibers can never go to true, fully b lack extinction under XP due to inherent , random ness in the structure of the cell. Consequently two examiners with slightly different interpretation s of the position of minimum brightness could in theory insert the compensator to perform the Herzog test and see mostly magenta with a hint of either orange or blue , making it difficult to conclude in which direction the c ellulose twists . Fortunately, this potential difficulty can usually be overcome in practice by rotating the stage to observe the ultimate a t other positions to make the colors more apparent and/or by observing multiple ultimates from the same sample to determine if the fiber has an S or Z twist. However, i n the initial observations, this phenomenon was thought to be linked to the species be ing examined. As the overall color of the Herzog test is determined by the S or Z direction of spiraling, it was proposed that the angle of the cellulose spiraling might be responsible for the observed differences in the Herzog test, namely that one specie s resulted in distinct blue and orange colors while the other resulted indistinguishable shades of magenta . 3.5 Raman spectroscopy of C. capsularis and C. olitorius ultimate cell wall s Several methods , including two dimensional X - ray diffraction ( 38 - 40 ) , scanning electron microscopy (SEM) ( 32 ) , and Raman spectr oscopy ( 33 - 35 ) , are available to assess the angle of cellul ose spiraling . However, both of these approaches have limitations which make them unamenable to adoption by forensic laboratories. First, t wo dimensional X - ray diffractometers are not widely available and the current financial environment in many laborator ies would prohibit the purchase of such powerful yet expensive instruments for such a narrow purpose . The 32 second method which held promise was utilizing SEM to directly observe the cellulose microfibrils and use trigonometry to calculate a spiraling angle ( 32 ) . This would have involved removing the outer layers of the cell wall to selectively expose the S2 layer of the secondary cell wall ( 32 ) . However, the publication presenting the SEM photomicrographs of wood fibers prepared in this manner did not include information about how to prepare the samples for such analysis ( 32 ) and attempts to re ach the author we re not successful . It remains to be seen whether this method for sample preparation could be reproduced. Thus, X - ray diffraction and SEM were not pursued in this work for the precise deter mination of the cellulose spiraling angle. Consequently, a nalysis b y Raman spectro scop y was selected to investigate the spiraling angle as it would entail the least expensive and simplest sample preparation. Raman spectroscopy is becoming more widely available in forensic laboratories, meaning its application to vegetable fibers could be adopted on a large scale. Sample preparation and analysis were available in the literature ( 34 ) and described in Section 1.7 . In that study by Gierlinger et al. , the authors used ratios of specific peak heights and partial least squares regression modeling of entire spectra in order to predict a spiraling angle ( 34 ) . The predicted angles were then verified for accuracy by comparison to angles determined by X - ray diffraction ( 34 ) . By modifying the comparison of peak height ratios, the work herein sought to determine if there could be a simpler method to indicate diffe rences in the spiraling angle. The peaks chosen for analysis were 1096 cm - 1 and 2983 cm - 1 . The former corresponds to the symmetric stretches of C - C, C - O, and C - O - C bonds oriented parallel to the direction of the cellulose while the later corresponds to t he symmetric C - H stretch perpendicular to the cellulose fibrils. The atoms involved in these critical bonds, highlighting how they align parallel and 33 perpendicular to the cellulose chain, are emphasized in the structure of cellulose appearing in Figure 11 . For this exploratory analysis, four Raman spectra were collected from the middle of the cell wall of a single cell from one reference specimen each of species. The ratio of the 1096 cm - 1 and 2983 cm - 1 peak heig hts was 1.81 for C. capsularis ( Figure 1 2 A ) and 1.76 for C. olitorius ( Figure 1 2 B ). Given the similarity of these ratios, the error introdu ced by manual baseline corrections, and the small sample size, this simplified peak height ratio method provided insufficient information to determine whether the cellulose spiraling angles could be similar or different between the samples. This method had been attempted due to the ease of calculating peak height ratios as opposed to using more complex statistical modeling of spectra to predict a precise angle. Taking advantage of such a shortcut could have made it more likely for such a technique to be ado pted by forensic laboratories owing to it being easier to understand the method, thereby reducing the time and effort needed to train analysts on the new method. However, it seems apparent that the full procedure described by Gierlinger et al . ( 34 ) would Figure 11. Structure of cellulose. The 1096 cm - 1 Raman band corresponds to the symmetric stretches of C - C , C - O, and C - O - C bonds oriented parallel to the cellulose microfibril; the oxygen atoms involved are colored blue for emphasis. The 2983 cm - 1 Raman band corresponds to the symmetric C - H bond oriented perpendicular to the cellulose microfibril; the invol ved hydrogen atoms are colored red for emphasis. 34 0 1000 2000 3000 4000 5000 0 500 1000 1500 2000 2500 3000 3500 Raman Intensity (CCD counts) Wavenumber (cm - 1 ) 0 500 1000 1500 2000 2500 0 500 1000 1500 2000 2500 3000 3500 Raman Intensity (CCD counts) Wavenumber (cm - 1 ) Figure 12. Average Raman spectra of C. capsularis and C. olitorius . Spectra were collected from 100 3,500 cm - 1 at four randomly chosen locations in the cell wall interior from a single ultimate from technical fiber cross sections. Spectra were baseline co rrected and zeroed before being averaged. A . One ultimate of C. capsularis (Sample B - 300 5 8) had a 1096 cm - 1 : 2983 cm - 1 height ratio of 1.81. B . One ultimate of C. olitorious (Sample B - 30059) had a ratio of 1.76. B . A . - - - - 35 indeed be necessary for predicting a cellulose spiraling angle that could be used to differentiate whether there are small differences in the cellulose spiraling angle between the two jute species. Thus, it remains undetermined whether there may be a diffe rence in the angle that has not yet been discovered or whether there is no difference in the angle. 3.6 New c onsiderations for the Herzog test The initial observations of the clarity or ambiguity of the colors observed during the Herzog test of Corchorus ultimates was initially thought to be tied to each species. This hypothesis, that the two species have a slightly different angle at which cellulose spiraled in the S2 secondary cell wall layer, drove the project early on to attempt to character ize this angle using Raman spectroscopy. After several more months of microscopically examining ultimates, however , a new hypothesis was developed , the theory of which is discussed below and visually summarized in Figure 13 . In isotropic fibers where n equals n , the material is not birefringent and thus is not visible (appears black) under XP ( 61 , 65 , 66 ) . This occurs for fibers with no regular internal arrangement of molecules; an example is fiberglass, as glass is a well - known amorphous solid ( 22 ) . Upon inserting the compensator, usually only an outline of the material is visible while its interior is the same magenta color as the field of view. In fibers such as polyester, the regular, semi - crystalline arrangement of polymer chains means the fiber is anisotropic ( 49 ) . In such fibers, n does not equal n and complete extinction occurs at the horizontal and vertical positions when viewed under XP. However jute fibers , which are anisotropic, do not go to complete extinction when oriented in the vertical and horizontal po sitions due to random variation in t he orientation of the 36 cellulose chains. In general, the darker the interference colors observed under XP, the lower the birefringence and the more simil ar the refractive indices of the fiber parallel ( n ) and Figure 13 . A new hypothesis to explain why some ultimates produce clear colors and others produce ambiguous colors during the Herzog test . At left, ultimates which appear brighter at extinction between XP produce clear orange or b lue colors during the Herzog test. This is predicted to be due to a more regular ordering or cellulose microfibrils in the cell wall. At right, ultimates which go to more complete extinction between XP result in the more ambiguous magenta color during the Herzog test. This is hypothesized to be due to more disorder in the orientation of cellulose microfibrils in the cell wall. 37 perpendicular ( n ) to the long axis of the fiber. These fibers that appear darker at extinction were observed to be the same ones that appear more magenta when viewed under XP with the compensator, producing an ambiguous result for the Herzog test ( Figure 13 ). In contrast, the jute ultimates that remain brighter and go to a less complete extinction are the same ones that appear more clearly orange in the horizontal position and blue in the vertical p osition ( Figure 13 ). Thus, it is hypothesized that ultimates displaying the more ambiguous, magenta color in the Herzog test which also appeared darker at extinction have more disorder in the orientation of their cellulose chains. Conversely the ultimates displaying clear oranges and blues in the Herzog test which were brighter at extinction are predicted to have a more ordered arrangement of cellulose microfibrils in the cell wall. SEM, X - ray diffraction, or Raman spectroscopy could be used to support thes e hypotheses if undertaken on a large scale for both species of Corchorus as well as other vegetable fibers. As much of the reviewed literature looked at small sample sizes for each species studied ( 33 - 35 , 38 , 40 ) , a larger investigation could provide a new understanding of whether the colors observe d in the Herzog test relate to the cellulose spiraling angle and, if so, how much the colors and angle varies in different plant species. 38 CHAPTER 4 : Results and Discussion Part 2: Microscopic analysis of commercial jute goods 4 . 1 Rationale for the analysis of commercial goods analysis Regardless of the persistent difficulties in distinguishing between fibers from each of the jute species, the ability to differentiate jute from other fiber types is necessary for more than just routine fiber identification in casework. Experienced examiners have noted that natural fiber products can contain fiber blends even if the products were advertised as a single fiber ( 53 ) . To illustrate, a poorly executed proficiency test from the 1990s required fiber analys ts to correctly identify manila hemp (also known as abaca, scientific name Musa textilis ) in a sample of unlabeled cordage. However, the cordage purchased and furnished by the testing company was in fact was compos ed of a mixture of manila and sisal (scientific name Agave sisalana ) , even though the label indicated it was only manila . Analysts who correctly identified both were penalized while those who incorrectly only identified manila received full marks ( 53 ) . Such proficiency tests impact the ability of analysts to maintain certification ( 67 ) and laboratories to maintain accreditation ( 68 ) . Not only does this blunde r illustrated why it is necessary to verify that product labels are accurate, it also demonstrates how vegetable fiber analysis is not as cut - and - dry as some may think. 4.2 Acquisition and summary of commercial goods One hundred and thirteen c ommercial goods containing or made from jute, or samples of such, were collected. A subset of local brick - and - mortar and online retailers provided the goods either by sale or donation. Th e criterion to acquire a sample for the commercial goods collection was whethe Some samples advertised as 39 products designed as substitutes for true jute burlap. The items included in the commercial goods collection are summarized in Table A.1 of Appendix A , which includes the unique sample identifier, the sample category, a description of the item, the source of the item, and the results of the analysis. All samples belonged to one of six categories: textiles, geotextiles, cordage, flooring, sacking, or miscellaneo us. Of the 113 commercial goods analyzed, 93 contained jute and 20 contained non - jute fibers ( Table 2 ) . These results will be reviewed in more detail in Sections 4.4 4.9 after a discussion of what characteristics were used to identify the fibers present in the commercial goods collection. 4.3 Features used to identify jute and non - jute fibers Representative fibers from each item were microscopically analyzed to determine if they contained jute or substitute fibers based primarily on the presence of chara cteristic features of jute as described in Sections 3.1 3.4 . This approach was chosen over others presented here Table 2 . S umma ry of commercial goods analysis by product type. Product type Total Number containing jute fibers Number containing other materials Textile 85 69 15 polyester 1 unverified polyester Geotextile 7 7 - Cordage 7 7 - Flooring 6 5 1 without fibrous material Sacking 6 3 1 plastic weave 2 undetermined vegetable fibers Miscellaneous 2 2 - Total 113 93 20 40 because it is recommended by the Scientific Working Group for Materials Analysis (SWGMAT) ( 44 ) and is well - aligned with current methods implemented by trace evidence analysts . Several other features not previously discussed were used to verify whether or not a fiber was jute. First, a drop or two of phloroglucinol reagent was applied to sample cuttings to assess the relative amount of lignin in the vegetable fibers. Highly ligni fied fibers are stained a bright or dark magenta hue and partially lignified fibers a light to medium pink, while fibers with little to no lignin do not change color ( 20 , 21 ) . The results of the phloroglu cinol test for selected reference vegetable fibers including jute, some jute substitutes, an d other common vegetable Figure 14. Photograph of the results of the phloroglucinol test for relative degree of lignification for reference samples of jute, some ju te substitutes, and some common vegetable fibers . Darker pink staining indicated a higher degree of lignification. Scale as indicated in the photograph. 41 fibers were photographed, as shown in Figure 14 and summarized in Table 3 . C. capsularis and C. olitorius stained a vibrant magenta, as expected. Agave sisalana and Cannabis sativa both stai ned more darkly than predicted. The thickness of the A. sisalana fibers may explain why the stain appeared darker than expected. Most commercial C. sativa products are softer than the reference sample used for this experiment; it is possible that this particular sample contained more lignin than is typical due to inco mplete extraction of the fiber from the plant or less processing than is typical for C. sativa commercial goods. In contrast, Musa textilis stained lighter than expected, although the very fine fibers of this sample may distort the perception of the color , making it appear paler . Linum usitatissimum had no visible color change as was expected. Finally, Hibiscus sabdariffa was expected to stain darker than intermediate pink but not quite as deeply magenta as jute. However, all three samples stained a bright magenta. The color by eye may have appeared slightly lighter than the deep magenta of the Corchorus samples, but not to an extent that this method could reliably distinguished these fibers. It is important to note that the phloroglucinol test is not specific for jute, as many fibers can exhibit a high degree of lignification. However, these results can support an identification of a fiber when observed in in conjunction with other features. T o this end, phloroglucinol was Table 3 . Summary of the results of the phloroglucinol test for relative degree of lignification for reference samples depicted in Figure 14. Sample number Species Expected color Observed color B - 30024 Corchorus capsularis Red to red - violet Magenta B - 30027 Corchorus olitorius Red to red - violet Magenta B - 30011 Agave sisalana Intermediate pink Magenta B - 30042 Musa textilis Intermediate pink Pale pink B - 30027 Linum usitatissimum No change to pale pink No change B - 30023 Cannabis sativa No change to pale pink Pale pink to magenta B - 30004 B - 30035 B - 30061 Hibiscus sabdariffa Lighter than jute Magenta (all) 42 added to all natural fibers in the commercial goods collection and the degree of staining was compared relative to the staining of the reference fibers noted above . The results are included in Table B.1 of A ppendix B . Most go ods reacted strongly and stained a deep magenta to indicate heavy lignification, which supported the conclusion of most goods as containing jute fibers . Exceptions included textiles that were artificially dyed such dark colors (e.g. black or deep purple) t hat it was not possible to observe a color change. A few also displayed an intermediate pink color, although this happened most commonly on very lightly colored fibers. These samples may have been bleached to achieve such light coloring. Because these fibe rs contained all other features of jute, this single observation was not deemed to be inconsistent with identifying these few samples as jute. Other macroscopics feature used to assist in the identification of jute from among other vegetable fibers is its characteristic heavy odor, texture when handled manually, and often earthy brown color. In general, bast fibers are typically softer than leaf fibers, and this holds true for jute when it is compared to fibers such as sisal. The nature of most jute goods obtained for this study is such that they are often more loosely woven, softer, and shed more fibers than other similarly available products. For example, sisal cordage from a hardware store may shed just as much as jute, but sisal has a more yellow color, is stiffer, and is less pungent. While hemp cordage might be of similar color and texture, it appears to shed slightly less and also has a characteristic odor that is easily distinguishable from that of jute. Such properties of gross samples can either fa cilitate an initial identification to later be confirmed or corroborate identification of a fiber based on microscopic characteristics . B ecause the amount of fiber evidence from a crime scene may be limited to only a single technical fiber, these properties are not always apparent for comparison. Nonetheless, experience with properties of bulk samples of 43 different vegetable fibers can provide an a nalyst with a valuable reference for comparison should larger samples be encountered in their casework, ensuring that no helpful information is overlooked. Finally, one of the most helpful features in the identification of jute is its characteristically v ariable lumen ( 17 , 27 ) . For the most part, reference samples from the Microtrace Vegetable Collection had lumen which varied from slightly more than half the overall diameter at the widest points to tightly pinched as the narrowest point s (refer to the thin and bolded arrows in Figures 5 and 6 ) . Further, this feature was observed in the vast majority of the commercial jute samples ( Figure 15 ) . However, some samples, especially many of the commercial samples, also had ultimates with less v ariable lumens. In these ultimates, the lumen occupied roughly one - third of the overall diameter at the maximum, therefore being slightly narrower than the typical jute ultimate lumen ( Figure 15 ) . These lumens were less frequently pinched at their narrowes t points and overall demonstrated more uniformity in diameter than the traditionall y telltale ultimates with widely varying lumens. Interestingly, these more uniform lumens occurred fairly frequently in commercial goods but were less frequently observed in reference samples. This initially caused some doubt in considering whether the commercial goods could be true jute, given the mismatch in how frequently the different lumen types were observed. However, other reputable sources suggest that the wider varia bility in lumen morphology in the commercial goods compared to the reference samples does not necessarily indicate that the commercial goods were not true jute. Despite finding the thinner, more uniform lumens infrequently in reference samples observed in this work, Catling and Greyson include photomicrographs of the lumens of reference C. capsularis and C. olitorius specimens ( 27 ) . These clearly depict not only a wide variety in relative lumen diameter within a single ultimate 44 but also provide proof that the thinner lumen does indeed occur in both species. Thus, the variation in lumen diameter observed in the commercial samples was concluded to fall within the range of expected variation for both Corchorus species. This assisted in preventing false exclusions due to the lack of thinner lumen in the reference samples observed for this project. 4.4 Analysis of textile samples Of the 85 textile samples, 78 swatches were obtained from two home decorating retailers and one special ty retailer, explaining why this product class is disproportionately represented in Figure 1 5 . Ultimates from a sample of jute flooring (Sample BF - 012) to demonstrate the variability in lumen morphology observed in many of the commercial goods. Solid arrows point of areas of ultimates demonstrating widely varying lumen diameter. Dashed arrows point to an ultimate demonstrating the more uniform and slightly narrower lumen morphology. Ulti mates were mounted in glycerin jelly and photographed under plane polarized light (PPL) at 100X total magnification. 45 t he overall collection of goods. Sixty - nine samples were verified to contain jute , 1 5 contained . One of the aforementioned home decorating retailers p rovided 10 samples, five composed of jute (BF - 016 through BF - 020) and five composed of man - made fiber s which were smooth to the touch (BF - 021 through BF - 025) . These latter five samples were identified as polyester on the basis of microscopically observed morphologica l and optical properties. Figure s 16 and 17 illustrate these features in Sample s BF - 02 3 and B F - 024 respectively , which were representative of this group of samples . All five swatches contained a blend of polyester fibers with an d without delusterant partic les that were in all other aspects similar ( Figure 16 ) . A ll Figure 1 6 . Blend of polyester fibers from a swatch of fuchsia textile advertised as - 023). Only the more lightly colored fibers contained lig ht - scattering delusterant particles. The relatively uniform diameter was indicative of a circular cross section. Fibers were mounted in Cargille oil of refractive index 1.70 and photographed under plane polarized light (PPL) at 200X total magnification. 46 circular cross - section as determined by the uniform diameter and symmetric pattern of birefringence colors under XP ( Figure 17 B ). Refractive indices for these five samples determined an n of approximately 1.70 and an n of approximately 1.53 to 1.55. From these refractive indices the calculated value for birefringence is approximately 0.15 to 0.17 , indicating a positive sign of elongation which is corroborated by the presence of addition colors in the northeast - southwest (NE - SW) position and subtractive colors in the northwest - southeast ( NW - Figure 1 7 . Photomicrograph series of a polyester fiber from a textile swatch advertised as - 024). A. Reference photomicrograph in PPL in the NE - SW position. B. High (possibly fourth) order interference colors were observed between XP. C. In the NE - SW position the ultimate exhibits addition of wavelengths into higher order pastel colors, approaching white, between XP with the compensator in place. D. In the NW - SE position the ultimate exhibits subtraction of wavelengths to lower order pinks. Together C and D illustrate that this fiber had a positive sign of elongation. The fiber was mounted in glycerin jell y and photographed at 100X total magnification. A. C. B. D. 47 SE ) position when the fibers are viewed under XP with the first order red compensator ( Figure s 17 C and 17 D ) . All of these characteristics are consistent with polyester. The web page listing all product samples, from which swatches were typically ordered, included the fiber composition for all other products except these ( 69 ) . Interestingly, this main webpage did not elaborate on the fiber co A customer could click to request more information, at which point individual pop - up boxes for these five products did disclose that they were composed of polyester. Because these five samples were less c learly labeled than as all other products, such fabrics could be ordered under the incorrect assumption that the product is dy ed burlap made from true jute. Curiously , the five samples from the same company that contain ed jute but were not advertised in th e product name as such. This information was again only found by again searching the detailed product description. This perhaps suggest that the visual appearance and texture of burlap is more desirable for interior decorating purposes, which is reasonable as true jute products shed substantially and may therefore not be ideal for many customers. The burlap specialty retailer furnished every sample in its collection for this study, a total of 62 samples. Fifty - eight were confirmed to be true jute while the remaining four ( BF - 026 through BF - 028 and BF - 032 ) were determined to be polyester on the basis of properties similar to those described earlier in this section . ( 70 ) , t he specifications for the jute burlap mention ed slight imperfections in the fabric, its biodegradability, a nd a warning that machine washing and drying can unravel and damage the fabric. In contrast, the four same rustic atmosphere as burlap ( 71 ) . Another online retailer 48 marketed their products (primarily table linens for events) as superior to true jute. The website noted shedding of fibers, the distinctive earthy odor, the inability to be machine washed, and the coarse texture as disadvantages when compared to the polyester material from which all their products were manufactured ( 72 ) . Similarly, another interior decorating company provided seven samples, six of which were polyester but only one of these was expressly advertised as faux burlap ( 73 ) . These advertisements from sellers may provide insight as to why polyester is being used by multiple companies to replace jute burlap; namely, that polyester is more easily cared for, less easily damaged , and longer lasting. These advantages likely explain the use of polyester as a man - made substitute fiber for jute, particularly for interior decorating purposes. 4.5 Analysis of geotextile samples Seven g eotextile samples , all consistent with true jute, were obtained from two individuals ass ociated with Michigan State University (MSU) Department of Horticulture. Thus they not retailers but were end users of the burlap geotextile s . None of the samples ( samples BF - 001 through BF - 003 and BF - 106 through BF - 109) c ould be attributed to a specific m anufacturer but all were leftovers from the usual course of their work for the Department of Horticulture. One provider stated that the burlap was purchased for wrapping tree roots while the other added that the burlap could also be used to prevent the ero sion of ecological features such as river banks. All were verified as true jute on the basis of macroscopic observation of the color, texture and odor of the bulk item; deep magenta staining in the phloroglucinol test indicating high lignin content; and mi croscopic observation of ultimates. 49 4.6 Analysis of cordage samples Six samples of cordage were obtained from a popular home improvement store, a dollar store, a Midwestern big - box store, and an online Ebay seller (samples BF - 010, BF - 089, BF - 098 through BF - 101 , and BF - 113 ). All were verified as consistent with true jute as described above. 4 .7 Analysis of flooring samples Five samples (BF - 011 through BF - 015) of different jute floor coverings were obtained from an online store specializing in vegetable fiber rugs. These five samples represented all of jute offerings and came with a rubbery backing to prevent skidding . All were found to contain jute fibers on the basis of the same macroscopic characteristics, reaction in the phloroglucinol test for lignin, and microscopic char acteristics as described above. A single sample of carpeting was obtained from a popular home improvement store upon asking if the store sold any carpet with jute backing. The employee was unsure if there was jute in the sample provided for this study, and indeed a visual examination determined that the backing was composed of a plastic mesh and no fibrous material w as apparent. This agreed with reports from c arpet manufacturers and the International Jute Study Group (IJSG) that the popularity of jute carpet backings has been steadily declining in favor of more durable synthetic materials ( 41 , 74 ) . 4.8 Analysis of sacking material samples Five samples of sacking materials were obtained for analysis. One (BF - 004) was excluded as it was provided by the MSU Department of Horticulture as an example of the synthetic materials replacing burlap sacking for sandbags. It was woven from strips of a p lastic 50 polymer material but was otherwise not fibrous in nature. The other five sacks were obtaine d from a local coffee retailer. As an end recipient rather than original merchant of the sacks, the shop was therefore not responsible for the advertising of the sack as jute or any other material. However, the staff was knowledgeable enough to provide three samples of sacks they believed t o be burlap and two sacks of a lighter - hued fiber which they could not identify. On the preliminary inspection, the three s amples (BF - 007 through BF - 009) had the characteristic brown color, scratchy but soft texture, and odor of true jute. These three sacks were used for the transportation of coffee beans from India, Indonesia, and Ethiopia. One sack (BF - 007) contained a tag i ndicating that the bag itself, as opposed to the coffee beans, originated from a jute mill in India. All criteria for the determination of the bags as jute we re met for these three samples. The other two sacks (BF - 005 and BF - 006) were used for the transpo rtation of decaffeinated green coffee beans from Colombia. These were a pale straw color, much scratchier than jute, less pliable than jute, and not as soft as jute . These features are all consistent with sisal ( Agave sisalana ) . Microscopic analysis reveal ed the presence of two fibers types ( Figure 18 ). The first had a very wide lumen, a diamond - shaped pattern along the entire ultimate, and lacked nodes, dislocations, and cross markings. The diamond pattern appears or original from dark markings that look like four - pointed stars. In general, these ultimates were somewhat shorter and plumper than jute ultimates. The star shapes and diamond pattern are similar to features of coir ( Cocus nucifera ) fiber ultimates, which most resembled the unknown fiber. Howeve r, a sample of coir fiber was not available for analysis and thus a definitive comparison of all features could not be made. The second, more common fiber in the sample had a more uniform lumen roughly one - third to one - half of the overall fiber diameter, few nodes and dislocations, and rare cross 51 Figure 1 8 . Blend of macerated fibers from coffee bean sacking (Sample BF - 005) and comparison to macerated coir fibers (Sample B - 30002). A. Numbered arrows point to the two types of fibers found in the macerated sample from the coffee sack. Fibers were mounted in glycerin jelly and photographed under PPL at 100X total magnification. B. Reference image of coir fibers to demonstrate similarity t o the first unknown fiber in the coffee sack. Fibers were mounted in water and photographed under PPL at 200X total magnification. A. B . 1 1 2 2 52 markings. This fiber demonstrated first order pale gray interference colors, had a positive sign of elongation , and twisted in the Z direction. Due to the similarity of the bulk sample with other macroscopic properties of sisal, this fiber was tentatively identified as sisal. When ashed, A. sisalana samples typically reveal a large number of acicular (needle - shaped) crystals o ccurring singly, in pairs, or in small groups ( 27 ) . Sometimes the crystals also appear curved like small black bananas if any leaf material remained attached to the crystals at the time of ashing ( 27 ) . While ashing samples was not possible with the equipment available in the laboratory, this method could have been used to confirm that A. sisalana was the primary fi ber present in the two coffee sacks. 4.9 Analysis of miscellaneous samples Two items in this category were obtained. The first (BF - 103) was a sample of fibers tweezed from a multi - level cat scratching post and climbing tower. The tag on the item indicate d that the fabric covering each layer of the climbing toward was jute. The second (BF - 112) was burlap twine trim around the edges of a decorative holiday ornament. Both were confirmed to be consistent with jute by the same methods as described for other sa mples. 4 . 10 T rends in jute fiber substitution in commercial goods That the geotextile, cordage, and miscellaneous jute produ cts were verified as true jute was unsurprising . The goods were marketed as such, and in general, a consumer expects a product lab el to be accurate. While 20 out of 113 examined jute or burlap products, or 18%, were found to contain non - jute materials , none of these 20 samples seemed to represent deliberate inaccuracies in 53 product labeling. All 16 of the non - jute textile samples were obtained from online retailers and durability, ease of maintenance, and burlap - like appearance of polyester likely ex plains why this synthetic fiber was found to be widely advertised as a substitute for true jute burlap, particularly for indoor applications. Additionally, the fact that a home improvement store employee advised that the store might no longer carry carpet with jute backing concurred with the IJSG statistics that few carpets are currently being manufactured with jute backings when more durable synthetic options are available. In contrast, it may initially be surprising that as sacking rose from 39% to 58% o f jute exports from India and Bangladesh in the last two decades , only half of the sacks analyzed in this study did not contain jute ( 41 ) . Certainly the sm all number of sacks analyzed limits any widely generalizable conclusions. However, sacking traditionally used for animal feed has largely been replaced with more durable synthetic materials ( 75 ) . The internationa l transportation of coffee beans evidently still utilizes jute sacks , as three were obtained for this project from a local coffee shop. Although two of the five coffee sacks were composed of non - jute fibers, it would be hasty to draw conclusions about how commonly each sack may be in this market. It is interesting to note that the non - jute vegetable fiber sacks were used solely for the transportation of decaffeinated coffee beans, perhaps as a way to provide packaging which prevents confusion between caffei nated and non - caffeinated varieties. Without knowing how much of the coffee bean market is occupied by caffeinated versus decaffeinated beans that adhere to such a packing scheme, it is not realistic to assume only half of jute sacks are actually jute. It must be remembered that these sacks were obtained from end - users rather than the entities that 54 manufacture, advertise, and sell them, and thus it may be that the two non - jute sacks would never have been mistaken for jute in the first place. 4. 11 Difficult ies encountered in the analysis of commercial goods Commercial goods typically showed slightly more variability in features than the reference samples. This may be possible due to the larger number of jute commercial samples (nearly 100) compared to the n umber of reference samples (nine). With regards to the lumen diameter, o may be due to the age of the plants when the fibers were harvested. The age of the samples affects ho w much of the secondary cell wall is deposited, with more of the wall being laid down over time . Plants that were older at the time of harvest have more time to lay down secondary cell wall than plants harvested earlier . It may be possible that the referen ce samples were collected from less mature jute plants than those used for the commercial goods. Further, it is unknown whether the manufacturers of jute goods typically use jute harvested from a single source or whether bales of jute from multiple sources are mixed together before using them to manufacture retail goods. Hence it is possible that the ultima tes observed from commercial goods more accurately represent lumen morphology of the entire jute population. While a country of origin and year of collection is available for all reference jute samples, it is impossible to definitively show if plant age at harvest might explain the wider variety in lumen morphologies among the commercial samples versus the reference samples. Another limitation of th is work was the use of characteristic fiber odors and textures to aid in identification. These are both inherently subjective criteria, which imply that the analyst must have sufficient experience with handling fiber samples to accurate interpret the signi ficance 55 of these two properties. If such mastery can be achieved, these properties may be helpful in ruling out jute from other fibers. For example, hemp ( C. sativa ) and flax ( L . usitatissitmum ), and sisal ( A. sisalana ) can be quickly eliminated from consi deration in this way. This can be particularly true if the ultimates of the two species are very similar to each other, as in the case of several jute substitutes, including roselle ( Hibiscus sabdariffa ), kenaf ( H. cannabins ), and aramina ( Urena lobata ). T o extend the utility of this approach, unsuccessful efforts were made to obtain any size reference sample of textiles made from these latter three fibers. The difficulty in obtaining swatches or any product made from these fibers may be interpreted as an i ndication of how rarely they are available in the United States. Thus, it was considered unlikely that any of these jute substitute fibers might have been used in any of the commercial goods. While odor and texture aided the identification of sample s as ju te, such interpretations should be made cautiously and not be used as a sole criterion by which to exclude the presence of other fibers in a sample. A final limitation of this work arises from bias introduced in which stores were selected in order to requ est or purchase samples. On a broad scale this study is limited to retailers willing to provide samples at low or no cost. Ideally, multiple stores of the same type (e.g. multiple home improvement stores) in multiple locations would have been visited. Furt her, more than one unit of each product would have been obtained in the same visit and by visiting the store over the course of several months or years. These measures would help identify if products may only occasionally contain non - jute fibers as a resul t of occasional changes in suppliers or represent more consistent substitutions. Such a rigorous sampling procedure would permit more general conclusions to be drawn about fraud trends in jute and burlap product mislabeling. 56 4.12 Photography of commercial goods samples for the future development of a jute fiber database All commercial goods were photographed macroscopically to record the physical state of the products at the time of purchase, sampling, or analysis. Additionally, a series of photomicrographs has been nearly completed for every sample. In this context, a complete set of photographs for a sample includes the macroscopic image of the sample and nine photomicrographs of one or more representatives ultimates oriented in four positi ons: the NE - SW position (one in PPL, one between XP, and one between XP with the compensator inserted), the NW - SE position, vertical, and horizontal (the latter all being collected both between XP and between XP with the compensator). This full r ange of im ages provides an invaluable, comprehensive repository of reference images which can be used to impartially confirm observations traditionally recorded in writing . Many highly - regarded reference texts suffer from a finite space in which to publish images, r esulting in inclusion of only one or two photomicrographs which cannot fully depict all microscopic features of a fiber ( 22 , 27 , 47 - 50 , 61 , 63 ) . This visual reference library intends to improve the uniformity of vegetable fiber analysis among fiber analysts across the country. New analysts can consult this compendium and evaluate for themselves morphological and optical phenomena uniq ue to different samples. Misinterpretation of the written observations and inappropriate a pplications of such information by novice analysts can become something of the past. The short - term goal is to electronically publish these images, along with the co rresponding written descriptions and interpretations, to serve as a freely available reference to forensic or other scientists interested in the natural variation in jute fibers and current trends in jute fiber substitutions. In addition, a summary of the frequency with which substitute fibers are 57 used in different classes of jute products will also be included with the database. The long - term goal is develop the database so that it can be updated with submissions by users of jute goods that they encounter in casework. This will serve the purpose of improving the scope of the goods included in the database to be more representative of the occurrence of jute goods available on the market. To date, no studies have examined how frequently different types of jut e goods are subject to misleading labeling or outright substitution of fibers. In laboratories with limited time and resources, this complication aims to be a useful commentary on current trends to assist examiners in a more thorough examination of particu lar product types. 58 CHA PTER 5 : Conclusions and Future Work 5.1 Efforts to identify a method to discriminate jute fibers from C. capsularis and C. olitorius Microscopic observations of morphology and optical properties, m easurement of ultimate dimensions , and simplified Raman analysis of the cell wall cellulose orientation were unable to differentiate ultimates from C. capsularis and C. olitorius . Without a rapid, inexpensive, and simple method to determine which species of jute is present in a sample it is in turn not possible to determine whether commercial jute goods contained one or both jute species. Such a method could therefore provide an additional level of discriminatory information about possible sources of questioned samples from crime scenes after initial identifying a sample as jute. Specifically, such information could aid an investigation if an item of jute evidence originated from only one species and known samples for comparison contained jut e from the same, the other, or both jut e species. The experiments detailed herein describe one of the first dedicated attempts to distinguish ultimates of the jute species in an effort to provide this additional information to forensic laboratories and investigators. A limitation of this appro ach is that it is currently unknown whether commercial goods often contain jute fibers from just one or both Corchorus species. Given that they have slightly different preferred habitats ( 13 , 14 ) , it may be that jute plants are harvested on site and only sent to nearby distributors with the result that manufacturing companies buying locally may, knowingly or unknowingly, be only using fibers from a single species of jute in their goods. Conversely, it is also possible that jute plants and fiber are sourced from such wide geographical regions that some or even all manufacturers produce goods which are a blend of fibers from bo th species. In the absence of a simple, rapid, and inexpensive technique for differentiating fibers of 59 the two species or a consistently traceable system of marking the origin of plants used in manufactured goods, most modern laboratories remain unable to determine which species are present in commercial available jute products. However, this need not be the end of the story. The Raman spectroscopy analysis performed in this work was a vastly simplified version of the analysis undertaken by the original au thors of the method ( 33 - 35 ) . Full statistical mode ling of Raman spectra from many samples of the two species may indeed reveal differences in the cellulose spiraling angle that the simplified analysis was unable to detect. Moreover, as Raman spectroscopy becomes more widely used for forensic cases ( 76 , 77 ) , increasing the number of applications of this technique improves the affordability of the instrument and serves to make it more a more widely accepted technique both in the laboratory and in the courts. As a result, it is suggested that any future efforts toward characterizing the cellulose spiraling angle of the jute fibers be focused on more closely reproducing the technique as it was applied by the original authors ( 33 - 35 ) . 5.2 Evaluating the frequency with which goods advertised to contain jute are composed of other fibers Slightly fewer than 18% of commercial textile, geotextile, cordage, flooring, sacking, and miscellaneous jute goods were determined to contain fibers other than jute. The overall lack of non - jute vegetable fibers was somewhat surprising, given the proficiency test mishap from the 1990s ( which partially inspired this project) and published literature on the frequency of natural fibers products being swapped or mislabeled ( 12 , 78 ) . However, some of the referring literature was published several decade s ago, likely before the widespread adoption of synthetic materials for such applications as livestock feed and carpet backing ( 41 , 74 , 75 ) . Such a decline in the use 60 of jute for these applications may have resulted in lesser demand, reducing former instances of substituting it out for less expensive or more readily available fibers. Furthermore, many of the jute text ile samples analyzed here were obtained from craft or specialty retailers catering to a developed world market with disposable income . T herefore it may not be surprising that polyester, a more durable fiber that can create fabrics with appearance of jute w ithout the inconveniences (shedding fibers, pungent earthy odors, and an inability to machine wash the fabric) , is currently the most common fiber substituted in goods marketed as jute. In general, the small number of samples analyzed for all product cat egories except textiles raises concerns about whether this analysis can be representative of all jute goods currently available in the United States and in the world. A limitation of this pilot study was that the collection of commercial goods was limited to what was inexpensively available in a limited geographical area over a short period of time and what was available inexpensively from online retailers . It follows that future studies could improve upon this work by including a more representative sample of jute products available on the market by obtaining multiple units of products from all stores selling these goods. This would help determine if some stores are more likely to carry mislabeled products. Additionally, repeatedly obtaining these goods fro m the same retailers would allow analysis of whether the frequency of mislabeling at a given store is relatively constant or if it changes over time, perhaps due to switches in suppliers. Detecting or reducing instances of fraud may be avoided by requiring shipments or batches of jute and vegetable fiber goods sold in the United States to provide certificates of verification clearly stating where the jute used in the item came from. Such protections may be difficult to enact and enforce, as much of the manu facturing takes place oversea s , even if it does ensure that the customer receives the product they are paying for. 61 By continuing to pursue the development of an online database of jute good s , the ability to accommodate user submissions of jute items enco u ntered in casework strives to improve its representation of jute products in circulation with time . Prior to this work, no studies examined how frequently jute mislabeling occurred in different types of products claiming to contain jute. Without this syste matic study of how often and in which product types such substitutions are likely to occur, the forensic fiber examiner is ill - equipped to know how likely it is that he or she is dealing with a jute substitute rather than true jute. Familiarity with the pr oducts in which substitutes commonly replace jute can thus guide an examiner toward a more thorough examination of particular products, thereby ensuring an efficient use of time and resources. 62 APPENDI X 63 Table A.1 . Description of items in commercial goods collection and results of microscopic analysis. Sample identifier Sample class Physical Condition Item description Source Analysis results BF - 001 Geo - textile Cutting Large, loosely woven burlap geotextile used for erosion control. Dan Bulkowski (MSU Horticulture) Jute BF - 002 Geo - textile Large piece Medium weave burlap for wrapping tree roots (horticulture). Medium - sized piece. May be from same source as BF - 003. Dan Bulkowski (MSU Horticulture) Jute BF - 003 Geo - textile Large piece Medium weave burlap for wrapping tree roots (ho rticulture). Large piece. May be from same source as BF - 002. Dan Bulkowski (MSU Horticulture) Jute BF - 004 Sacking Large piece Synthetic burlap. Often used for sandbags. Dan Bulkowski (MSU Horticulture) Woven plastic polymer BF - 005 Sacking Whole Sacking used for transporting decaf green coffee beans from Colombia, South America. Some red and green threads used. Bag printed with information. Front: A. LAUMAYER, GREEN COFFEE BEANS, 70 kg Net, 3 02 0458. A red number "20" is written and circled. Reverse: ARM ENIA EXCELSO, PRODUCT OF COLOMBIA, 3 02 0458. The Coffee Barrel 2237 Aurelius Rd. Holt, MI http://www.thecoffeebarrel.com/index.phtml Unidenti - fied vegetable fiber 64 Table A.1 ( cont'd) . BF - 006 Sacking Whole Sacking used for transporting decaf green coffee beans from Colombia, South America. Some red and green threads used. Bag printed with information. Front: A. LAUMAYER, GREEN COFFEE BEANS, 70 kg Net, 3 02 0458. Reverse: ARMENIA EXCELSO, PRODUCT OF COLOMBIA, 3 02 0458. The Coffee Barrel 2237 Aurelius Rd. Holt, MI http://www.thecoffeebarrel.com/index.phtml Unidenti - fied vegetable fiber BF - 007 Sacking Whole Sacking used for transporting coffee beans from India. Bag printed with information. Front: INDIAN COFFEE, MONSOONED MALABARAA, ALLANA Reverse: C:111400000633, 14/53/2012/38/C.No.26738, OAKLAND, UNITED STATES, NETT 50 KILOS, 233 There is also a tag on the bag itself. The tag reads: FOOD GRADE JUTE BAG, MADE IN INDIA, HOWRAH JUTE MILL, H5JM(illegible letter)NC3QL The Coffee Barrel 2237 Aurelius Rd. Holt, MI http://www.thecoffeebarrel.com/index.phtml Jute 65 Table A.1 ( cont'd) . BF - 008 Sacking Whole Sacking used for transporting coffee beans from Ethiopia. One strand of hot pink fiber used. Bag printed with information. Front: BAGERSH, PRODUCE OF ETHIOPIA, 010/0006/0028, CERT.NO.0028. Handwritten: 20210. Reverse: WAS HED, YIRGACHEFFE, GRADE 2, ORGANIC, BCS OKO GARANTIE The Coffee Barrel 2237 Aurelius Rd. Holt, MI http://www.thecoffeebarrel.com/index.phtml Jute BF - 009 Sacking Whole Sacking used for transporting coffee beans from Indonesia. Several trans of teal fibers used. Bag printed with information. Front: 015/8828 (illegible), JAVA EST (illegible), WASHED ARABICA GRADE 1, WONOSOBO, PROD. OF INDONESIA. The Coffee Barrel 2237 Aurelius Rd. Holt, MI http://www.thecoffeebarrel.com/index.phtml Jute BF - 010 Cord - age Whol e Three pack of 36 m jute twine. Two spools are natural color, one speel is green. Label states the brand is Tool Bench Hardware. Made in China, imported into the US by Greenbrier International, Inc. at 500 Volvo Parkway, Chesapeake, VA 23320. Dollar Tree, 5823 West Saginaw Highway, Lansing, MI 48917 Jute 66 Table A.1 ( cont'd) . BF - 011 Floor - ing Swatch 8cm x 8.5cm swatch. Rug swatch with rubber - like backing to improve adhesion to floor to reduce slippage. Sample name: Jute Mahal, White. Color is ivory. Some dark spots. Sisal Rug Direct online store Order #100033338 Contact: sales@sisalrugs.com Website: http://www.sisalrugs.com/ Jute BF - 012 Floor - ing Swatch 8cm x 8.5cm swatch. Rug swatch with rubber - like backing to improve adhesion to floor to reduce slippage. Sample name: Jute Mahal, Birch. Color is uniform, cool, light brown. Sisal Rug Direct online store Order #100033338 Contact: sales@sisalrugs.com Website: http://ww w.sisalrugs.com/ Jute BF - 013 Floor - ing Swatch 8cm x 8.5cm swatch. Rug swatch with rubber - like backing to improve adhesion to floor to reduce slippage. Sample name: Jute Mahal, Artic Gold. Multi - colored from ivory to a warm, honey reddish - brown. Sisal Rug Direct online store Order #100033338 Contact: sales@sisalrugs.com Website: http://www.sisalrugs.com/ Jute BF - 014 Floor - ing Swatch 8cm x 8.5cm swatch. Rug swatch with rubber - like backing to improve adhesion to floor to reduce slippage. Sample name: Jute Ma hal, Wheat. Color is light, warm brown. Sisal Rug Direct online store Order #100033338 Contact: sales@sisalrugs.com Website: http://www.sisalrugs.com/ Jute BF - 015 Floor - ing Swatch 8cm x 8.5cm swatch. Rug swatch with rubber - like backing to improve adhesion to floor to reduce slippage. Sample name: Jute Mahal, Clay. Color is light, warm brown. Sisal Rug Direct online store Order #100033338 Contact: sales@sisalrugs.com Website: http://www.sisalrugs.com/ Jute 67 Table A.1 ( cont'd) . BF - 016 Textile Swatch 13cm x 14cm swatch. Sample name: Meridian Woodsmoke (SID: 592) Natural color with print. Regal Drapes online store Website: www.regaldrapes.com Jute BF - 017 Textile Swatc h 13cm x 14cm swatch. Sample name: Meridian White (SID: 591) Ivory color with print. Regal Drapes online store Website: www.regaldrapes.com Jute BF - 018 Textile Swatch 13cm x 14cm swatch. Sample name: Meridian Natural (SID: 590) Gray with print. Regal Drapes online store Website: www.regaldrapes.com Jute BF - 019 Textile Swatch 13cm x 14cm swatch. Sample name: Lumine White (SID: 545) Ivory with blue (silky, non - jute) embroidery. Regal Drapes online store Website: www.regaldrapes.com Jute BF - 020 Textile Swatch 13cm x 14cm swatch. Sample name: Lumine Fall Leaf (SID: 544) Gray with black (silky, non - jute) embroidery. Regal Drapes online store Website: www.regaldrapes.com Jute BF - 021 Textile Swatch 13cm x 14cm swatch. Sample name: Vintage Burlap Khaki (SID: 867) Regal Drapes online store Website: www.regaldrapes.com Polyester BF - 022 Textile Swatch 13cm x 14cm swatch. Sample name: Vintage Burlap Orange (SID: 869) Regal Drapes online store Website: www.regaldrapes.com Polyester BF - 023 Textile Swatch 13cm x 14cm swatch. Sample name: Vintage Burlap Fuchsia (SID: 865) Regal Drapes online store Website: www.regaldrapes.com Polyester BF - 024 Textile Swatch 13cm x 14cm swatch. Sample name: Vintage Burlap Ivory (SID: 866) Regal Drapes online store Website: www.rega ldrapes.com Polyester 68 Table A.1 ( cont'd) . BF - 025 Textile Swatch 13cm x 14cm swatch. Sample name: Vintage Burlap Avocado (SID: 862) Regal Drapes online store Website: www.regaldrapes.com Polyester BF - 026 Textile Swatch Faux Burlap Collection 60" roll 100% polyester Color: Vintage Jute (cool, light variegated brown) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Poly ester BF - 027 Textile Swatch Faux Burlap Collection 60" roll 100% polyester Color: Cinnamon (warm, medium brown) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Polyester BF - 028 Textile Swatch Faux Burlap Collection 60" roll 100% polyester Color: Winter Wheat (light, neutral wheat brown) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Po lyester BF - 029 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Bridal Rose The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 030 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Ceremony White The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 031 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Espresso The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 032 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Winter Wheat (Faux) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Polyester 69 Table A.1 ( cont'd) . BF - 033 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Perfect Pink The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jut e BF - 034 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Rainforest The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 035 Textile Swatch The Bridal & Event Collection 11 oz/48" roll Color: Sea Glass Green The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 036 Textile Swatch 11 oz/60" burlap Color: Natur al The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 037 Textile Swatch 11 oz/60" burlap Color: Barrel Brown The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 038 Textile Swatch 11 oz/60" burlap Color: Antique Lace The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http:// www.theburlapfactory.com/ Jute BF - 039 Textile Swatch 11 oz/60" burlap Color: Espresso Bean The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 70 Table A.1 ( cont'd) . BF - 040 Textile Swatch 11 oz/60" burlap Color: Desert Sand The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website : http://www.theburlapfactory.com/ Jute BF - 041 Textile Swatch 11 oz/60" burlap Color: Terra Cotta The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 042 Textile Swatch 11 oz/60" burlap Color: Antique Brass The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 043 Textile Swatch 11 oz/60" burlap Color: Hazel The Burlap Factory/ Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 044 Textile Swatch 11 oz/60" burlap Color: Rust The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 045 Textile Swatch 11 oz/60" burlap Color: Pumpkin The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 046 Textile Swatch 11 oz/60" burlap Color: Golden The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 71 Table A.1 ( cont'd) . BF - 047 Textile Swatch 11 oz/60" burlap Color: Wheat The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 048 Textile Swatch 11 oz/60" burlap Color: Raspberry The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 049 Textile Swatch 11 oz/60" burlap Color: Craftsman Green The Burlap Fa ctory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 050 Textile Swatch 11 oz/60" burlap Color: Hampton Blue The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 051 Textile Swatch 11 oz/60" burlap Color: Midnight The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 052 Textile Swatch 11 oz/60" burlap Color: Camo (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 053 Textile Swa tch 11 oz/60" burlap Color: Chevron (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 72 Table A.1 ( cont'd) . BF - 054 Textile Swatch Holiday Collection Color: Vintage Plaid (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 055 Textile Swatch Holiday Collection Color: Shimmer Ivory & Gold (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 056 Textile Swatch Holiday Collection Color: Shimmer Natural & Gold (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 057 Textile Swatch Holiday Collection Color: Shimmer Red & Gold (10 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 058 Textile Swatch Holiday Collection Color: Pure Red (11 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 059 Textile Swatch Holiday Collection Color: Eucaliptus (11 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 060 Textile Swatch Holiday Collection Color: Evergreen (11 oz) The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 73 Table A.1 ( cont'd) . BF - 061 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Crisp White The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 062 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Corn Silk The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 063 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Natural The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 064 Te xtile Swatch 11 oz/48" roll Shalimar Burlap Color: Farm Table The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 065 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Willow The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 066 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Storm The Burlap Factory/Big Duck Canvas 7 41 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 067 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Granit Grey The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 74 Table A.1 ( cont'd) . BF - 068 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Jet Black The Burlap Fa ctory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 069 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: True Red The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 070 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Ribbon Pink The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Web site: http://www.theburlapfactory.com/ Jute BF - 071 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Apricot The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 072 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Copper The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 073 Textile Swatch 11 oz/48" roll Shalimar Bur lap Color: Amethyst The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 074 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Spirt Orange The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 75 Table A.1 ( cont'd) . BF - 075 Textile Swatch 11 o z/48" roll Shalimar Burlap Color: Sour Apple The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 076 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Avocado The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 077 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Evergreen The Burlap Factory/Big Duck Canvas 741 West Winde r Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 078 Textile Swatch 11 oz/48" roll Shalimar Burlap Color: Eucalyptus The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 079 Textile Swatch 22 oz/48" Color: Natural The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 080 Textile Swatch 10 oz Color: Peacock Blue The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 081 Textile Swatch 10 oz Color: Hunt Club Green The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute 76 Table A.1 ( cont'd) . BF - 082 Textile Swatch 10 oz Color: Primrose Pink The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 083 Textile Swatch Color: Black The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkw ay Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 084 Textile Swatch 10 oz/60" Color: Wheat The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 085 Textile Swatch 10 oz Color: French Vanilla The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 086 Textile Swatch 10 oz Color: Light Olive The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 087 Textile Swatch 10 oz Color: Deep Red The Burlap Factory/Big Duck Canvas 741 West Winder Industrial Parkway Winder, GA 30680 - 7807 Website: http://www.theburlapfactory.com/ Jute BF - 088 Textile Swatch Faux Burlap The Burlap Shop online store 1050 Northfield Ct. Suite 300 Roswell, GA 30076 Website: www.theburlapshop.com Phone: 770 - 442 - 8777 E - mail: Sales@TheBurlapShop.com Pol yester 77 Table A.1 ( cont'd) . BF - 089 Cord - age Large piece Welt Cord Jute #10 5/32" Sold by the yard/meter Ebay seller: northwesttarpandcanvas Northwest Tarp & Canvas, LLC Jim Watt (Owner) Onsite manufacturing, boat canvas repair, heavy duty tarps, custom covers 703 West Holly Street Bellingham, WA 98225 Jute BF - 090 Textile Swatch Upholstery supplies fabric Jute webbing No strip for crafts, wedding 3 3/4" Ebay seller w holesale_upholstery_supplies Shipped from Stephen Sickles PO Box 142 Beecher Falls VT 05902 Shipped with a leather sample stamped with the image of a steer head and the words "Dave Robertson, morethanleader.com, Steve Stickles" Jute BF - 091 Textile Swatch Burlap www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Jute BF - 092 Textile Swatch Faux burlap www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Polyester (unverified) 78 Table A.1 ( cont'd) . BF - 093 Textile Swatch Jute 100% polyester Black www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Polyester BF - 094 Textile Swatch Jute 100% polyester Gray www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 P olyester BF - 095 Textile Swatch Jute 100% polyester Natural www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Polyester BF - 096 Textile Swatch Jute 100% polyester Slate Blue www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Polyester BF - 097 Textile Swatch Jute 100% polyester White www.brightsettings.com P.O. Box 374 701 East Spring Street Titusville, PA 16354 Phone: 866 - 827 - 4177 Fax: 866 - 827 - 7747 Polyester 79 Table A.1 ( cont'd) . BF - 098 Cord - age Whole Mibro KingCord Heavy Duty Jute, 190 feet, twisted Natural color Manufactured to MIBRO's exacting specifications in China The MIBRO Group, Buffalo, NY 14225; Toronto, Ontario M1L 4S6 www.mibro.com 302071 0 66366 30207 3 LA302071FR - D Meijer retail store #25 2055 W. Grand River Ave Okemos, MI 48864 Jute BF - 099 Cord - age W hole Mibro KingCord Fine Jute, 190 feet, twisted Green color Manufactured to MIBRO's exacting specifications in China The MIBRO Group, Buffalo, NY 14225; Toronto, Ontario M1L 4S6 www.mibro.com 301511 0 66366 30151 9 3 LA301511FR - D Meijer retail store #25 2 055 W. Grand River Ave Okemos, MI 48864 Jute BF - 100 Cord - age Whole 100% Jute twine, #30 x 190 ft Natural color 7 lb working load limit Distributed by Crown Bolt, Aliso Viejo, CA 92656 Made in China 0 30699 65325 6 172540 The Home Depot 1749 Newman Rd Okemos, MI 48864 Jute 80 Table A.1 ( cont'd) . BF - 101 Cord - age Whole 100% Garden Jute Twine, #30 x 200 ft Green color 7 lb working load limit Made in China Distributed b y Home Depot, 2455 Paces Ferrt Rd NW, Atlanta, GA 30339 0 30699 12795 5 172540 SKU 494 250 12795 The Home Depot 1749 Newman Rd Okemos, MI 48864 Jute BF - 102 Floor - ing Swatch Shaw Style/Color: Viking Buckwheat - THS Construction: 100% BCF Olefin Product Part: 0701649725 Order SKU: 326 - 686 Sample SKU: 629 - 742 Note: Store worker said he had heard the middle layer (between the olefin fibers and plastic backing) is made of jute, but couldn't find any documentation to corroborate that. The Home Depot 1749 Newman Rd Okemos, MI 48864 No fibrous material present 81 Table A.1 ( cont'd) . BF - 103 Miscel - laneous Loose fibers Loose fibers tweezed from a jute and paper multi - level condo cat furniture. Fibers tweezed only from jute level coverings, not paper rope around the posts. Tag front reads: "51 in. after assembly, paper rope & jute construction, 5 scratching post & perch, 4 levels of fun" Tag reverse reads: "51 in x 22 in x 15 in, KEY 062, 3394964, 7 08820 39834 4, Dist. by Meijer Distribution, Inc. Grand Rapids, MI 49544, Made in China, www.meijer.com" Meijer retail store #25 2055 W. Grand River Ave Okemos, MI 48864 Jute BF - 104 Textile Loose fibers Loose fibers tweezed from t he burlap lining of a decorate basket. Tag front reads: "The Rococo Collection, Burlap Lined wire Basket, 12.6 in x 8.7 in x 4.3 in, whitmor" Tag reverse reads "6708 - 5630, 0 38861 63182 8, Made in China, (C)2014 Whitmor, Inc., 8680 Swinnea Road Suit 105, S outhaven, MS 38671, 1 - 888 - WHITMOR, help@whitmor.com, www.whitmor.com" Meijer retail store #25 2055 W. Grand River Ave Okemos, MI 48864 Jute 82 Table A.1 ( cont'd) . BF - 105 Textile Loose fibers Loose fibers tweezed from a burlap canvas type soft crate. Tag front reads: "Home store" Tag reverse reads: "Natural Burlap Crate, 15 in L x 10 in W x 11.5 in H, 3281253, 838, 8 86926 45627 1, Dist. By Wholesale Merchandisers LL C, 2929 Walker, NW, Grand Rapids, MI 49544, Made in China, Nuerrh, DNS1002" Tag inside product reads: "Shell: 100% burlap exclusive of decoration, spot clean only, made in China" Meijer retail store #25 2055 W. Grand River Ave Okemos, MI 48864 Jute BF - 106 Geo - textile Large piece 36" x 36" untreated burlap sheet Product 3636UR Professor Tom Fernandez, MSU Horticulture Burlap from A.M. Leonard (www.amleo.com) Jute BF - 107 Geo - textile Large piece 48" x 48" untreated burlap sheets Product 4848UR Professor Tom Fernandez, MSU Horticulture Burlap from A.M. Leonard (www.amleo.com) Jute BF - 108 Geo - textile Whole Burlap sock, old, with holes, source uncertain Professor Tom Fernandez, MSU Horticulture Burlap from A.M. Leonard (www.amleo.com) Jute BF - 109 Geo - textile Cutting Burlap sock cutting, medium golden colored Professor Tom Fernandez, MSU Horticulture Burlap from A.M. Leonard (www.amleo.com) Jute 83 Table A.1 ( cont'd) . BF - 11 2 Miscel - laneous Whole Wooden fox - shaped Christmas ornament lined with burlap ModCloth online retailer Jute BF - 113 Cord - age Cutting FD107 10M 3 - ply twisted burlap string natural ribbon fiber jute twine rope toy L Ebay seller: leisure - z Jute BF - 115 Textile Swatch Bengal Burlap Natural #BLAP. 100% jute, 6.4 oz per square yard, 46/47" wide. Thread count is 11 threads per inch x 10 threads per inch. Dharma Trading Company Jute BF - 116 Textile Swatch Bengal Burlap Bleached #BLAPB. 100% jute, 6.4 oz per s quare yard, 46/47" wide. Thread count is 11 threads per inch x 10 threads per inch. Dharma Trading Company Jute 84 Table A . 2 . O bservations of fibers fr om the commercial goods collection. Sample identifier Analysis results Physical properties Optical properties Aggregate observations of all jute reference samples Not applicable Lumen: Diameter typically varied within a single ultimate from approximately half of overall diameter to tightly pinched. Some ultimates had a lumen diameter up to three - fourths of the overall diameter. Some ultimates had lumen diameters that were slightly thinn er and more uniform, ranging from approximately one - fifth to one - third of the overall diameter; these cells had at least one section of lumen diameter varying widely over a short distance. Texture: Irregular and occasionally rough. Some ultimates with wide r lumen appeared to have a texture similar to wood grain. Lignification: Stained bright or deep magenta upon application of phloroglucinol reagent, indicating a high degree of lignification Other cell types: Not necessarily present. Pitting: Not always obs erved. Odor: Characteristic, pungently earthy odor detectable in large samples Dislocations and nodes: Common and not evenly spaced. Cross markings: Occasional to extremely frequent and not evenly spaced. Interference colors: Pale, first order yellows to s econd order bright blues. Sign of elongation: Positive. Direction of cellulose twist determined by the Herzog test: Z. Non - birefringent crystal - like inclusions: Present in the sample but not necessarily in all ultimates. BF - 001 Jute Consistent with jute C onsistent with jute BF - 002 Jute Consistent with jute Consistent with jute BF - 003 Jute Consistent with jute Consistent with jute BF - 004 Woven plastic polymer Not analyzed Not analyzed 85 Table A. 2 ( cont'd) . BF - 005 Unconfirmed vegetable fibers. Fiber 1 was not able to be identified. Fiber 2 is similar to sisal ( Agave sisalana ). Two fibers were present in a blend Bulk sample observations Texture: Bulk sample much stiffer than jute. Lignification: Bright magenta indicative of highly lignified fibers Other cell types: Not noted Pitting: Not noted Odor: Characteristic, sharp odor of sisal was detectable in large samples. Color: Sacking was a pale, straw yellow color. Fiber 1 Lumen: Ver y wide. Texture: Diamond - shaped pattern along the entire ultimate. General: Most ultimates of this type are broken. In general the ultimates are shorter and plumper than jute or sisal ultimates. Fiber 2 Lumen: Roughly one - third to one - half the overall dia meter. Lumen could become pinched but in general was less variable than is typically observed in jute ultimates. Common to both fiber types Interference colors: First order grays Sign of elongation: Positive Non - birefringent crystal - like inclusions: Not no ted Fiber 1 Dislocations and nodes: Absent Cross markings: Absent Direction of cellulose twist determined by the Herzog test: Not recorded Other features: The diamond pattern visible in plane polarized light is accentuated. The pattern appears to originat e from four - pointed stars. The star shapes are superficially similar to but more prominent than those that appear in coir ( Cocos nucifera ) ultimates. Fiber 2 Dislocations and nodes: Few Cross markings: Rare Direction of cellulose twist determined by the Herzog test: Z 86 Table A. 2 ( cont'd) . BF - 006 Unconfirmed vegetable fibers. Fiber 1 was not able to be identified. Fiber 2 is similar to sisal ( Agave sisalana ). Two fibers were present in a b lend Bulk sample observations Texture: Bulk sample much stiffer than jute. Lignification: Bright magenta indicative of highly lignified fibers Other cell types: Not noted Pitting: Not noted Odor: Characteristic, sharp odor of sisal was detectable in larg e samples. Color: Sacking was a pale, straw yellow color. Fiber 1 Lumen: Very wide. Texture: Diamond - shaped pattern along the entire ultimate. General: Most ultimates of this type are broken. In general the ultimates are shorter and plumper than jute or s isal ultimates. Fiber 2 Lumen: Roughly one - third to one - half the overall diameter. Lumen could become pinched but in general was less variable than is typically observed in jute ultimates. Common to both fiber types Interference colors: First order grays Sign of elongation: Positive Non - birefringent crystal - like inclusions: Not noted Fiber 1 Dislocations and nodes: Absent Cross markings: Absent Direction of cellulose twist determined by the Herzog test: Not recorded Other features: The diamond pattern vis ible in plane polarized light is accentuated. The pattern appears to originate from four - pointed stars. The star shapes are superficially similar to but more prominent than those that appear in coir ( Cocos nucifera ) ultimates. Fiber 2 Dislocations and nod es: Few Cross markings: Rare Direction of cellulose twist determined by the Herzog test: Z BF - 007 Jute Consistent with jute Consistent with jute BF - 008 Jute Consistent with jute Consistent with jute BF - 009 Jute Consistent with jute Consistent with jute BF - 010 Jute Consistent with jute Consistent with jute BF - 011 Jute Consistent with jute Consistent with jute 87 Table A. 2 ( cont'd) . BF - 012 Jute Consistent with jute Consistent with jute BF - 013 Jute Consistent with jute Consistent with jute BF - 014 Jute Consistent with jute Consistent with jute BF - 015 Jute Consistent with jute Consistent with jute BF - 016 Jute Consistent with jute Consistent with jute BF - 017 Jute Consistent with jute Consistent with jute BF - 018 Jute Consistent with jute Consistent with jute BF - 019 Jute Consistent with jute Consistent with jute BF - 020 Jute Consistent with jute Consistent with jute BF - 021 Polyester Apparent cross section shape: Circular Diameter: Delusterant particles: Absent n : close to or slightly above 1.70 n : between 1.53 and 1.55 Birefringence: 0.15 to 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 022 Polyester Apparent cross section shape: Circular Diameter: Delusterant particles: Absent n : close to or slightly above 1.70 n : 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 023 Polyester Sample homogenous: No Apparent cross section shape : Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : close to or slightly above 1.53 Birefringence: 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive 88 Table A. 2 ( cont'd) . BF - 024 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : close to or slightly above 1.53 Birefringence: 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 025 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Prese nt n : 1.70 n : between 1.53 and 1.55 Birefringence: 0.15 to 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 026 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: Delusterant partic les: Present in some fibers and absent in others. n : close to or slightly greater than 1.70 n : between 1.53 and 1.55 Birefringence: 0.15 to 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 027 Polyester Sample homogeno us: No Apparent cross section shape: Circular Diameter: Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : between 1.53 and 1.55 Birefringence: 0.15 to 0.17 Interference colors: High (third - fourth) order Sign of e longation: Positive BF - 028 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 029 Jute Consistent with jute Consistent with jute 89 Table A. 2 ( cont'd) . BF - 030 Jute Consistent with jute Consistent with jute BF - 031 Jute Consistent with jute Consistent with jute BF - 032 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: Delusterant particles: Present in some fibers and absent in others. n : close to or slightly less than 1.70 n : close to or slightly greater than 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 033 Jute Consistent with jute Consistent with jute BF - 034 Jute Consistent with jute Consistent with jute BF - 035 Jute Consistent with jute Consistent with jute BF - 036 Jute Consistent with jute Consistent with jute BF - 037 Jute Consistent with jute Consistent with jute BF - 038 Jute Consistent with jute Consistent with jute BF - 039 Jute Consistent with jute Consistent with jute BF - 040 Jute Consistent with jute Consistent with jute BF - 041 Jute Consistent with jute Consistent with jute BF - 042 Jute Consistent with jute Consistent with jute BF - 043 Jute Consistent with jute Consistent with jute BF - 044 Jute Consistent with jute Consistent with jute BF - 045 Jute Consistent with jute Consistent with jute BF - 046 Jute Consistent with jute Consistent with jute BF - 047 Jute Consistent with jute Consistent with jute BF - 048 Jute Consistent with jute Consistent with jute BF - 049 Jute Consistent with jute Consistent with jute BF - 050 Jute Consistent with jute Consistent with jute BF - 051 Jute Consistent with jute Consistent with jute 90 Table A. 2 ( cont'd) . BF - 052 Jute Consistent with jute Consistent with jute BF - 053 Jute Consistent with jute Consistent with jute BF - 054 Jute Consistent with jute Consistent with jute BF - 055 Jute Consistent with jute Consistent with jute BF - 056 Jute Consistent with jute Consistent with jute BF - 057 Jute Consistent with jute Consistent with jute BF - 058 Jute Consistent with jute Consistent with jute BF - 059 Jute Consistent with jute Consistent with jute BF - 060 Jute Consistent with jute Consistent with jute BF - 061 Jute Consistent with jute Consistent with jute BF - 062 Jute Consistent with jute Consistent with jute BF - 063 Jute Consistent with jute Consistent with jute BF - 064 Jute Consistent with jute Consistent with jute BF - 065 Jute Consistent with jute Consistent with jute BF - 066 Jute Consistent with jute Consistent with jute BF - 067 Jute Consistent with jute Consistent with jute BF - 068 Jute Consistent with jute Consistent with jute BF - 069 Jute Consistent with jute Consistent with jute BF - 070 Jute Consistent with jute Consistent with jute BF - 071 Jute Consistent with jute Consistent with jute BF - 072 Jute Consistent with jute Consistent with jute BF - 073 Jute Consistent with jute Consistent with jute BF - 074 Jute Consistent with jute Consistent with jute BF - 075 Jute Consistent with jute Consistent with jute BF - 076 Jute Consistent with jute Consistent with jute BF - 077 Jute Consistent with jute Consistent with jute BF - 078 Jute Consistent with jute Consistent with jute 91 Table A. 2 ( cont'd) . BF - 079 Jute Consistent with jute Consistent with jute BF - 080 Jute Consistent with jute Consistent with jute BF - 081 Jute Consistent with jute Consistent with jute BF - 082 Jute Consistent with jute Consistent with jute BF - 083 Jute Consistent with jute Consistent with jute BF - 084 Jute Consistent with jute Consistent with jute BF - 085 Jute Consistent with jute Consistent with jute BF - 086 Jute Consistent with jute Consistent with jute BF - 087 Jute Consistent with jute Consistent with jute BF - 088 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : Close to or slightly greater than 1.55 Birefringen ce: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 089 Jute Consistent with jute Consistent with jute BF - 090 Jute Consistent with jute Consistent with jute BF - 091 Jute Consistent with jute Consistent with jute BF - 092 Polyester (unverified) Not analyzed Not analyzed BF - 093 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : 1.70 n : Close to or slightly greater than 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive 92 Table A. 2 ( cont'd) . BF - 094 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 12 - Delusterant particles: Present in some fibers and absent in others. n : close to or slightly less than 1.70 n : Close to or slightly less than 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) ord er Sign of elongation: Positive BF - 095 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : close to or slightly greater than 1.70 n : Close to o r slightly less than 1.55 Birefringence: 0.15 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 096 Polyester Sample homogenous: No Apparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : close to or slightly less than 1.70 n : Between1.53 and 1.55 Birefringence: 0.15 to 0.17 Interference colors: High (third - fourth) order Sign of elongation: Positive BF - 097 Polyester Sample homogenous: No A pparent cross section shape: Circular Diameter: 15 - Delusterant particles: Present in some fibers and absent in others. n : close to or slightly greater than 1.70 n : close to or slightly less than 1.55 Birefringence: 0.15 Interference colors: High ( third - fourth) order Sign of elongation: Positive BF - 098 Jute Consistent with jute Consistent with jute BF - 099 Jute Consistent with jute Consistent with jute BF - 100 Jute Consistent with jute Consistent with jute BF - 101 Jute Consistent with jute Consistent with jute BF - 102 No fibrous material present Not analyzed Not analyzed 93 Table A. 2 ( cont'd) . BF - 103 Jute Consistent with jute Consistent with jute BF - 104 Jute Consistent with jute Consistent with jute BF - 105 Jute Consistent with jute Consistent with jute BF - 106 Jute Consistent with jute Consistent with jute BF - 107 Jute Consistent with jute Consistent with jute BF - 108 Jute Consistent with jute Consistent with jute BF - 109 Jute Consistent with jute Consistent with jute BF - 112 Jute Consistent with jute Consistent with jute BF - 113 Jute Consistent with jute Consistent with jute BF - 115 Jute Consistent with jute Consistent with jute BF - 116 Jute Consistent with jute Consistent with jute 94 BIBLIOGRAPHY 95 BIBLIOGRAPHY 1. Kirk PL. Microscopic evidence: Its use in the investigation of crime. J Crim L & Criminology. 1949;40(3):362 - 9. 2. Grieve MC. The role of fibers in forensic science examinations. Journal of forensic sciences. 1983;28(4):877 - 87. 3. Petraco N. The occurrence of trace evidence in one examiner's casework. 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