Nature helps to solve the crime – Diatoms study in case of drowning death

Diatoms study in case of drowning death ISSN: 2394-0026 (P)Review Article ISSN: 2394-0034 (O)Nature helps to solve the crime – Diatoms study in case of drowning deathPragnesh Parmar1*, Gunvanti B. Rathod2, Sangita Rathod3, Ashish Parikh41Associate Professor, Forensic Medicine Department, SBKS MI & RC, Vadodara, Gujarat, India 2Assistant Professor, Pathology Department, SBKS MI & RC, Vadodara, Gujarat, India3Assistant Professor, Department of Medicine, AMC MET Medical College, Ahmedabad, India 4Consultant Physician, Gayatri Hospital, Gandhinagar, Gujarat, India *Corresponding author email: [email protected] to cite this article: Pragnesh Parmar, Gunvanti B. Rathod, Sangita Rathod, Ashish Parikh. Naturehelps to solve the crime – Diatoms study in case of drowning death. IAIM, 2014; 1(3): 58-65. Available online at www.iaimjournal.comReceived on: 11-10-2014 Accepted on: 25-10-2014AbstractNature is everywhere surrounding us in different form. It includes single cell structure to extremelycomplex, living or non living, substance or creature. One of the important part of nature is diatoms –unicellular algae which are usually present in water sources like lake, pond etc. Death due tosubmersion of body into water is known since ages. Diatoms study in case of drowning death is veryuseful for forensic investigators. It is one of the best example in which nature helps to solve thecrime. In present article, we discussed about diatoms, diagnosis of drowning death and length ofsubmersion of body in water with help of diatoms, collection, identification and preservation ofdiatoms for forensic analysis.Key wordsNature, Drowning, Diatoms, Crime, Submersion.Introduction which are usually present in water sources like lake, pond etc. Death due to submersion of bodyNature includes single cell structure to into water is known since ages. Diatoms study inextremely complex, living or non living, case of drowning death is very useful forsubstance or creature. One of the important forensic investigators. It is one of the bestpart of nature is diatoms – unicellular algae example in which nature helps to solve the crime. As typical features of ante-mortemInternational Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 58Copy right © 2014, IAIM, All Rights Reserved.

Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O)drowning disappear very rapidly with successfully utilized it to solve a case of acommencement of putrefaction, diatom test drowning mystery in 1904. The method of itshad gained the importance as gold standard for extraction was improved by acid digestion of thediagnosis and confirmation of drowning deaths. tissues. Incze successfully detected diatoms in blood and parenchymal organs in 1942 [2].Diatoms and other algae Tamasaka detected diatoms in bone marrow in 1949 [3]. In the 1960's and 1970's, TimpermanThe term ‘algae’ includes a wide range of using large series of drowning cases, providedorganisms varying from single celled protists to evidence for the validity of diatom test [4].colonial forms to huge multi cellular seaweedssuch as kelp. Almost all algae contain Otto (1961) reported on workers in Kieselguhrchloroplasts. Although algae are ubiquitous in industry who had contracted silicosis throughterrestrial and aquatic ecosystems, only the continuous inhalation of diatoms in the lungs,diatoms are useful in forensic analysis. Diatoms without of course, having drowned [5]. Spitzare unicellular algae belonging to the phylum (1963) and Peterson (1963) showed diatoms toStramenopila (phylum Bacillariophyta) and are be present in the major organs of non-drownedcharacterized by silica containing cell walls subjects, and came to conclusion that thecalled as ‘frustules’. They can be present in fresh diatom method was worthless [6, 7]. Porawskiwater and salt water, as well as the surface of (1966) also found diatoms in the lungs, kidneysmoist terrestrial habitats. Each frustule is made and bone marrow of non-drowned subjects [8].up of two halves, which are called as valves, one Koseki (1968) reported their occasional presenceof which is slightly smaller than the other so that in the lungs (and rarely in the liver and kidney)fit together one inside the other. Many species of non-drowned bodies [9]. Schellmann andof diatom have a characteristic frustule design Sperl (1979) had found diatoms in the bonewhich helps in their identification. There are marrow of 15 out of 16 non-drowned subjectsnumerous species of diatoms and more than [10].5600 have been described and probably morethan 1,00,000 in total. Their abundance and Tamaska (1949) found no diatoms in the bonespecies composition vary between locations marrow of seven subjects who were shot beforeowing to different diatoms preferring different entering the water [3]. Muller (1963) did notconditions such as temperature, salinity and pH find diatoms in the liver of 30 non-drownedand the consequences of inter - species bodies [11]. Neidhart and Greendyke (1966)competition. Therefore, the presence of failed to detect diatoms in the organs of 15 non-individual species or the species composition in drowned bodies [12]. Peabody AJ (1977), Ludesa forensic sample can provide an identifying B et al., (1994) and Pollanen MS (1997) stronglyfeature of a habitat or location at a particular supported diatom test in diagnosis of drowningtime of year. [13, 14, 22, 23]. Pollanen MS claimed that diatoms found in non-drowning cases couldHistorical aspects of diatoms in relation to have caused by contamination during variousdrowning process of autopsy and diatom test [22, 23]. Bortolotti F, et al., (2004) advocated the use ofDiatom was first detected in lung fluid by Environmental Scanning Electron MicroscopeHofmann E [1]. However, Rovenstorff had (ESEM) over light microscope for better diagnosis of diatom [15]. Wang Lei, et al., (2004)International Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 59Copy right © 2014, IAIM, All Rights Reserved.

Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O)advocated maximum value of diatom aspirated river and subsequently been swept out. Theinto the lung (number of diatoms per tissue in possibility that a person may have died in agram) to differentiate drowning and non- water body other than the one in which theydrowning deaths [16]. were found needs to be borne in mind when deciding on extraction techniques. For example,Diagnosis of drowning by help of diatoms the frustules of marine diatoms are dissolved by Soluene - 350 (a solubilizing agent) whilst theDrowning is the mechanical asphyxia death frustules of freshwater diatom species are notcaused by aspiration of fluid into air passages [20]. Like all biological evidence, the results ofdue to submersion of body (nose and mouth) diatom analysis need to be considered ininto water or fluid. Death due to drowning is context. For example, some workers questionvery complex and mechanism of death is the sensitivity of the diatom test and theasphyxia. Autopsy findigs of drowning very absence of diatoms does not mean drowning diddifficult because signs of immersion only not occur, nor does the presence of diatoms inindicate that a body was underwater (or some the body tissues mean that it did. Althoughother fluid) for certain time and not that the diatoms are extremely common, they are notperson died of drowning. Because diatoms can found in all water sources and even if they werebe found in many water sources, their presence present they may not find their way into thein the lungs and other tissues has been used as body organs or be recovered if they did.an indication that the victim may have drowned Similarly, the abundance of diatoms means that[17]. Diatoms can be recovered from the contamination at the time the body wasstomach (indicating that water was swallowed), retrieved, during the autopsy or duringfrom the lungs (indicating that it was aspirated) laboratory analysis is always a possibility unlessand also from the blood, major organs and the extreme care is taken. Furthermore, if a personbone marrow (indicating the water was repeatedly swims in a lake or the sea, it isaspirated whilst the victim was still alive). Owing possible that they may accumulate diatomsto their small size and the damage caused to the within their tissues over time so finding themlungs during drowning, diatoms may pass does not mean that the person drowned therethrough the alveoli and be swept around the [21].body in the blood stream [18]. Diatoms may also be recovered from the tissuesHowever, once the blood circulation stop after of persons who do not die of drowning or swimdeath, any diatoms entering the lungs with regularly. Diatoms are found in numerous manwater would not be transported elsewhere. By made products ranging from building materialscomparing the abundance and species profile of to the powder used in rubber gloves. There arethe diatoms found in the body with that of therefore many opportunities in which diatomsdiatoms found in the river – or wherever the may be breathed in and there is also a possibilityvictim was recovered – it is possible to provide that they might be absorbed through thecorroborating evidence with results from the gastrointestinal tract when consumed withautopsy to determine whether or not they foodstuffs although the extent to which thisdrowned and if so, whether it was at that occurs needs to be confirmed by furtherlocation [19]. For example, the presence of research. Consequently, diatoms can be found infreshwater diatoms in a body recovered at sea the tissues of persons who died of causes otherwould suggest that the victim may have died in a than drowning. However, it is generally acceptedInternational Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 60Copy right © 2014, IAIM, All Rights Reserved.

Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O)that finding diatoms within the bone marrow Numerous types of algae, including diatoms,provides good corroborative evidence of display seasonal growth quality and some aredrowning and identification to species level can infamous for forming short lived toxic blooms.exclude those that are contaminants [22, 23]. Therefore, if an effort is to be done at comparing the species composition recoveredDetermination of length of submersion and from body and specific area of lake or pond, itexposure by growth of algae may be essential to take sequential samples through the year and, in the case of open water,Length of submersion can be estimated by rate samples should be taken from both the surfaceof colonization of submerged body by diatoms waters and the bed. In the case of a river orand other algae but there are fewer studies in stream, samples should also be taken boththis field [24]. Depth, substrate, local above and below the site at which the body wascircumstances and time of year cumulatively found because there may be differences in theaffect the amount of growth of algae and diatom flora and the body may have beentherefore experimentation will help for any moved by the water currents. Due to the smallprediction. For example, in nutritionally rich size of diatoms and their widespreadpond, growth would be rapid in summer than distribution, every try should be done to evadeduring winter and compare to in nutrient poor the possibility of sample contaminationmountain stream. Haefner, et al. (2004) occurring in both the field and the laboratory.described use of algal growth to determine the All collecting equipments must be cautiouslylength of time a dead body remained in water cleaned before use, and the samples should be[25]. Instead of identification of the species of processed in clean glassware within thealgae and their relative large quantity, they laboratory and laboratory procedures must bedetermined total algal density by measuring the adopted to lessen the chances of contamination.amount of the chlorophyll pigment in a sample Samples of distilled water should be processedcollected. at the same time to check contamination.Exposed objects and animal remains in Diatoms and other algae can be collected fromterrestrial environments can be colonized by open water source by using plankton nets whichalgae and their growth could be assessed. So consist of a long funnel shaped net bag mountedcalled ‘blue - green algae’ are not algae at all but on a circular frame and with a collecting vialthey are prokaryotic organisms because they attached to the narrower trailing end. The meshlack an enclosed nucleus and membrane - bound size of the net will determine the size of theorganelles and are more accurately known as plankton which is usually small and require acyanobacteria. In spite of being tremendously fine net size of 0.1 – 0.3 mm. Algae and othercommon in both global and aquatic ecosystems microscopic organisms attached to underwatertheir forensic potential has received little notice, substrates are referred to as ‘periphyton’ andalthough signs of colonization of exposed bones require specialized collection techniques [25].is said to become clear to the naked eye after 2 After collection, samples may need further– 3 weeks under suitable situation [26]. concentration, for example by centrifugation, although the method needs to be chosen withCollection, identification and preservation concern if the more delicate species of algae areof diatoms and algae for forensic analysis to be preserved undamaged. The samples can be examined directly using an ordinary stageInternational Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 61Copy right © 2014, IAIM, All Rights Reserved.

Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O)microscope although phase contrast illumination diatom species and found that the CO1improves the detail examination. If one is only (cytochrome oxidase subunit 1) gene offeredinterested in the diatoms and there is a lot of considerable potential [32]. This is somewhatcontaminating organic matter present, the surprising because although this gene issamples may be air dried and subjected to acid commonly used in phylogenetic studies ofdigestion although the majority of ordinary animals it is considered to have limitedalgae do not have silicaceous cell walls and applicability for plants. Abe, et al. (2003)would be destroyed. attempted to identify the presence of diatoms in forensic samples using molecular techniques butDiatoms can be extracted from body tissues, soil although they found the approach to beand other solid substances, by heating with sensitive and it could not be used to identifyconcentrated nitric acid for up to 48 hours [17, sufficient species of diatoms to distinguish the27]. Then after digest is centrifuged, the locality at which a person drowned [33]. Itsupernatant material is discarded, and the pellet should be remembered that molecularis washed by one or two cycles of suspension in techniques could not be used on diatomdistilled water and centrifugation after which specimens isolated from soil or tissues usingthe final pellet is microscopically examined. This standard extraction techniques since the strongmethod depends upon the silicaceous frustules acids would destroy their DNA. Anotherwhich remains after all organic matter is alternative identification technique is to usedissolved away but some authors like to use less Fourier transform infrared (FT - IR) spectroscopydangerous reagents and have adopted DNA to distinguish between species on the basis ofextraction technique [28] and ultrasonic their chemical composition. FT - IR has proveddigestion [29]. capable of distinguishing species that are difficult to tell apart on the basis of theirMorphological features of diatoms and algae morphology [34] and useful for populationhelp in their identification [30] but it is a time studies [35] but it does not appear to be inconsuming and often very difficult process. widespread use for identifying diatoms. EveryAlternately, number of other processes has been life has an end on one day by natural ordiscovered. Complex three dimensional shapes unnatural means. Death due to accident orof diatom frustules can be detected by drowning leaves a bad taste in society [36, 37,computer pattern recognition software and 38]. Study of nature in form of diatoms to solvethere have been future development of drowning death is very useful.automatic devices that can simultaneouslyidentify and count different diatom species [31]. SummaryHowever, even if automatic devices can beperfected, molecular evidence indicates that Nature is everywhere around us and helps us inmorphology on its own is a poor species many way, one of it is to solve crime. Diatomsindicator for diatoms and the existence of study in case of drowning death is one of the‘cryptic’ and ‘pseudo species’ is widespread. This best examples of it. Invention in the fields ofis an important consideration in a forensic physics, chemistry, biology and science helpscontext in which the identification of species human to use nature for different purposes.composition is to form part of the evidence. Effective use of nature without deteriorating it isEvans, et al. (2007) tested the suitability of a the best way to serve humankind.range of genes for phylogenetic analysis ofInternational Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 62Copy right © 2014, IAIM, All Rights Reserved.

Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O)Acknowledgement drowning by the diatom method. Acta Med Boil., 1968; 15: 207-19.Authors acknowledge the immense help 10. Schellmann B, Sperl W. Diotomeenreceived from the scholars whose articles are Nachweis im Knochenmark (femur)cited and included in references of this Nichtertrunkener. Z Rechtsmed., 1979;manuscript. The authors are also grateful to 83: 319-24.authors / editors /publishers of all those articles, 11. Mueller B. [on the problem of thejournals and books from where the literature for occurrence of diatoms in the organs ofthis article has been reviewed and discussed. cadavers not having lain in the water.] [Article in German - Original Title: ZyrReferences Frage des Vorkommens von Diatomeen in Organen von Leichen die nicht in 1. Hofmann E. Lehrbuch der Gerichtlichen Wasser gelegen haben] Dtsch Z Gesamte Medicin (\"Textbook of Legal Medicine\"). Gerichtl Med., 1963; 54: 267-72. 1878, p. 629. 12. Neidhart DA, Grrendyke RM. The significance of diatom demonstrations in 2. Incze G. Fremdkörper in Blutkreislauf the diagnosis of death by drowning. Am Ertrunkener. Zentralbl Allg Pathol Anat. J Clin Pathol., 1966; 48(4): 377-82. 1942; 79: 176. 13. Peabody AJ. Diatoms in Forensic Science. J Forensic Sci Soc., 1977; 17: 81- 3. Tamasaka L. Vizihullak Csontvelojneck 7. Diatomattartalmarol (Diatom Content of 14. Ludes B, Quantin S, Coste M, Mangin P. Bone Marrow in Corpses in water). Orv Application of simple enzymatic Hetil., 1949; 16: 509-11. digestion methods for diatom detection in the diagnosis of drowning in putrefied 4. Timperman J. The diagnosis of drowning corpses by diatom analysis. Int J Leg - a review. Journal of Forensic Science, Med., 1994; 107: 37-41. 1972; 1: 397-409. 15. Bortolotti F, Tagliaro F, Manetto G. Objective diagnosis of drowning by the 5. Otto H. uber den Nachweis von “diatom test”- A critical review. Forensic Diatomeen in menschlichen Sci Rev., 2004; 16: 135-148. Lungenstauben. Frankf Z Pathol., 1961; 16. Lei W, Jei W, Ying-kang H. Determination 71: 176-81. of maximum value of diatom aspirated into the lung and its practical application 6. Spitz WU, Schneider V. The significance in diagnosis of drowning cases. Journal of diatoms in the diagnosis of death by of Guiyang Medical College, 2004. drowning. J. Forensic Sci., 1964; 9(1): 11- 17. Pollanen M.S. Diatoms and homicide. 8. Forensic Science International, 1998; 91: 29–34. 7. Peterson F. Diatomeenbefunde bei 18. Lunetta P., et al. Scanning and Wasserleichen. Deutsch Ztschr Ger transmission electron microscopical Med., 1963; 54: 376-78. evidence of the capacity of diatoms to penetrate the alveo-capillary barrier in 8. Porawski F. Investigation on the occurrence of diatoms in organs in death from various causes. J For Med., 1963; 13: 134-87. 9. Koseki T. Fundamental examination of experimental materials and control animals on the diagnosis of death fromInternational Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 63Copy right © 2014, IAIM, All Rights Reserved.

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Diatoms study in case of drowning death ISSN: 2394-0026 (P) ISSN: 2394-0034 (O) tertiary care hospital of Ahmedabad, Gujarat. IAIM, 2014; 1(2): 17-22. Source of support: Nil38. Parmar P. Dowry death and law – Indian Conflict of interest: None declared. scenario. IAIM, 2014; 1(2): 44-49.International Archives of Integrated Medicine, Vol. 1, Issue. 3, November, 2014. Page 65Copy right © 2014, IAIM, All Rights Reserved.