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Differentiation at Autopsy Between in Vivo Gas Embolism and Putrefaction Using Gas Composition Analysis

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Differentiation at Autopsy Between in Vivo Gas Embolism and Putrefaction Using Gas Composition Analysis Int J Legal Med DOI 10.1007/s00414-012-0783-6 ORIGINAL ARTICLE Differentiation at autopsy between in vivo gas embolism and putrefaction using gas composition analysis Yara Bernaldo de Quirós & Oscar González-Díaz & Andreas Møllerløkken & Alf O. Brubakk & Astrid Hjelde & Pedro Saavedra & Antonio Fernández Received: 18 January 2012 /Accepted: 4 October 2012 # Springer-Verlag Berlin Heidelberg 2012 Abstract Gas embolism can arise from different causes New Zealand White Rabbits models: control or putrefaction, (iatrogenic accidents, criminal interventions, or diving relat- infused air embolism, and compression/decompression. The ed accidents). Gas analyses have been shown to be a valid purpose of this study was to look for qualitative and quan- technique to differentiate between putrefaction gases and titative differences among groups and to observe how pu- gas embolism. In this study, we performed systematic nec- trefaction gases mask in vivo gas embolism. We found that ropsies at different postmortem times in three experimental the infused air embolism and compression/decompression models had a similar gas composition prior to 27- – Electronic supplementary material The online version of this article h postmortem, being typically composed of around 70 – (doi:10.1007/s00414-012-0783-6) contains supplementary material, 80 % of N2 and 20 30 % of CO2, although unexpected which is available to authorized users. higher CO2 concentrations were found in some decom- : Y. Bernaldo de Quirós A. Fernández (*) pressed animals, putting in question the role of CO2 in Veterinary Histology and Pathology, Department of Morphology, decompression. All these samples were statistically and Institute of Animal Health, Veterinary School, significantly different from more decomposed samples. University of Las Palmas de Gran Canaria (ULPGC), Gas composition of samples from more decomposed ani- Trasmontaña s/n. 35416, Arucas, Las Palmas, Spain mals and from the putrefaction model presented hydrogen, e-mail: [email protected] which was therefore considered as a putrefaction marker. O. González-Díaz Keywords Putrefaction . Gas embolism . Decompression . Physical and Chemical Instrumental Center for the Development . of Applied Research Technology and Scientific estate (CIDIA), Gas composition Nitrogen Edificio Polivalente 1, University of Las Palmas de Gran Canaria (ULPGC), Campus de Tafira s/n, 35017, Introduction Las Palmas, Spain A. Møllerløkken : A. O. Brubakk : A. Hjelde Air embolism (AE) is the entry of atmospheric or alveolar air Department of Circulation and Medical Imaging, into the vascular system and is mainly an iatrogenic problem Norwegian University of Science and Technology, [1]. In many cases, it arises as a complication of numerous Trondheim, Norway invasive medical procedures [1], criminal intervention, or P. Saavedra barotraumas [2]. However, gas embolism of other origins Department of Mathematics, can occur as a consequence of significantly supersaturated University of Las Palmas de Gran Canaria (ULPGC), gas tissues after decompression [3], when the sum of the Campus de Tafira s/n, 35017, Las Palmas, Spain dissolved gas tensions (oxygen, CO2,nitrogen,andhelium) and water vapor exceeds the local absolute pressure [4]. Present Address: The diagnosis of gas embolism at autopsies is based on Y. Bernaldo de Quirós morphological findings and on the chemical analysis of the Biology Department, Woods Hole Oceanographic Institution, MS#50, gas collected from the heart [5]. To avoid atmospheric air Woods Hole, MA 02543, USA entrance as a result of dissection, Richter’s technique is Int J Legal Med typically used [6]. Gas chromatography can differentiate Experimental putrefaction model between gas embolism and putrefaction gases [2, 5, 7, 8]. Putrefaction is a continual process based on tissue break- Anesthetized NZWR were euthanized with an intraperitone- down by microorganisms such as bacteria, fungi, and pro- al injection of 200 mgkg−1 diluted pentobarbital. Euthanized tozoa (from the intestine and the environment) following animals were kept in hermetically sealed plastic boxes for autolysis [2] and results in the production of gases, liquids, biological material at room temperature (22.9–24.6 °C) for and simple molecules. Therefore, differentiation between 1, 3, 6, 12, 27, 42, 47, 53 (n01 for each time) and 67 h (n0 gas embolism and putrefaction gases might be postmortem 2) PM prior to necropsy. (PM) time and decomposition status-dependent. PM time is unknown on many occasions, and putrefaction processes are Experimental infused AE model dependent on the weather, especially temperature and hu- midity. Decomposition status can be coded based on the Anesthetized NZWR received atmospheric air infusion morphological status of the external and internal organs of through a catheter (0.36 mm I.D.) placed in the central vein the animal [9]. Since environmental conditions are expected of the ear with the use of a pump at 2.2 mLmin−1 until the to vary with different crime scenarios, comparison of gas rabbit expired. The total volume of air infused varied be- composition within the same decomposition code seems tween 4.5 and 13 mL. The dead animals were placed in more reliable. There are few systematic studies, using labo- hermetically sealed plastic boxes for biological material at ratory animals, that compare gas composition with PM time room temperature (23–25.3 °C) for 0, 20, and 40 min, and 1, or decomposition status [10, 11]. Such studies have tradi- 3, 6, 12, 27, 42, 53, and 67-hPM (n01 for each time). tionally focused on iatrogenic air embolism, while other gas embolism causes, such as those involving supersaturated Compression/decompression model tissues, after decompression have not been studied in much detail: In these cases, high nitrogen content is expected [12]. In this case, anesthetized NZWR were compressed in pairs in Although for diving-related accidents gas analysis is also a dry, hyperbaric chamber (Animal Chamber System, NUT, sometimes performed [13], there are no studies on how Haugesund, Norway) to eight absolute atmospheres during a these gases evolve with PM time or how they interact with bottom time of 45 min, followed by a fast decompression putrefaction gases. (0.33 ms−1). Diving profile was selected for explosive decom- In the present study, we analyzed gas composition in pression induction. Animals that died were placed in hermet- three rabbit experimental models (putrefaction, infused air ically sealed plastic boxes for biological material at room embolism, and gases produced by decompression) and its temperature (23–25 °C) for 0, 20, and 40 min, and 1, 3, 6, evolution with PM time and decomposition status. 12, 27, 42, 53, and 67-hPM (n02 for each time). Animals that survived for 1 h after decompression were euthanized with an intraperitoneal injection of 200 mgkg−1 diluted pentobarbital. Material and methods Only those animals that died because of decompression (n0 11) (without euthanasia) were considered for this study. Experimental procedures PM procedures A total of 41 New Zealand White Rabbits (NZWR, Animal Supply Center of the Negrín Hospital, Spain and the Norwe- Complete necropsies were carried out for each animal at its gian University of Science and Technology, Norway) of 2.5– scheduled PM time. In addition to PM time, a decomposition 3.8 kg were used. The animals were divided into three exper- code from 1 (very fresh) to 5 (advanced putrefaction) based on imental groups: (1) control, experimental putrefaction model the conservation state of the body was given to each animal (n010); (2) infused AE model (n011); and (3) compression/ [9]. Necropsy was performed with the rabbit in dorsal decu- decompression model (n020). All experiments were per- bitus position. Dissection was done carefully to avoid cutting formed in accordance with the European Union regulations of large vessels. First, the skin was removed and the abdom- for laboratory animals and were conducted under surgical inal cavity opened to expose the vena cava. Secondly, the anesthesia (Medetomidine of 0.5 mgkg−1 and Ketamine of thoracic cavity was opened, allowing access to the heart. 25 mgkg−1, subcutaneously). Experimental protocols for the Finally, the animals were completely submerged in water to infused AE model and for the putrefaction model were ap- sample the gas and to avoid atmospheric air pollution. Gas proved by the Ethical Committee for Animal Experiments of sampling, storage, and analysis were performed in accordance the University of Las Palmas de Gran Canaria (Spain). The to Bernaldo de Quirós et al. [14]. Gases were sampled from Norwegian Committee for Animal Experiments approved the the intestine, right heart, left heart, vena cava, and interstitial protocol for the compression/decompression model. emphysema where present. A total of 158 samples were Int J Legal Med th obtained (41 from the putrefaction model, 56 from the infused where Ni,H denotes the nitrogen observed in the i animal at AE model, and 61 from the compression/decompression mod- the Hhours postmortem, ai is the effect of the animal, and el). Gas composition of samples was calculated in micromoles finally, ei,H is the variability within each animal. Temporal and normalized in micromole percentage. Samples with an variation for gases of C/D was not detected in either cluster atmospheric air-like composition were considered as polluted during the considered period (β100), but it was detected in and were excluded from the study (n05). the AE model (β1<0). In
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