Performance of a Novel Electronic Nose for the Detection of Volatile Organic Compounds Relating to Starvation or Human Decomposition Post-Mass Disaster.

There has been a recent increase in the frequency of mass disaster events. Following these events, the rapid location of victims is paramount. Currently, the most reliable search method is scent detection dogs, which use their sense of smell to locate victims accurately and efficiently. Despite their efficacy, they have limited working times, can give false positive responses, and involve high costs. Therefore, alternative methods for detecting volatile compounds are needed, such as using electronic noses (e-noses). An e-nose named the 'NOS.E' was developed and has been used successfully to detect VOCs released from human remains in an open-air environment. However, the system's full capabilities are currently unknown, and therefore, this work aimed to evaluate the NOS.E to determine the efficacy of detection and expected sensor response. This was achieved using analytical standards representative of known human ante-mortem and decomposition VOCs. Standards were air diluted in Tedlar gas sampling bags and sampled using the NOS.E. This study concluded that the e-nose could detect and differentiate a range of VOCs prevalent in ante-mortem and decomposition VOC profiles, with an average LOD of 7.9 ppm, across a range of different chemical classes. The NOS.E was then utilized in a simulated mass disaster scenario using donated human cadavers, where the system showed a significant difference between the known human donor and control samples from day 3 post-mortem. Overall, the NOS.E was advantageous: the system had low detection limits while offering portability, shorter sampling times, and lower costs than dogs and benchtop analytical instruments.

Decomposition and insect succession on human cadavers in a humid, continental (Dfb) climate (Quebec, Canada).

Forensic entomology has been developing globally for decades. Despite this discipline being used in criminal investigations around the world, only a few controlled studies have been performed on human cadavers in human decomposition facilities, with the majority of these being conducted in warm and often dry climates. Therefore, the purpose of our research was to catalogue the first published data on insects associated with decomposed human bodies in a humid, continental (Dfb) climate. Specifically, our objective was to document the diversity and succession of the entomofauna associated with human cadavers throughout the decomposition process, in the Quebec province of Canada, during the summer season. Two human cadavers were studied in 2020 at the site for Research in Experimental and Social Thanatology, REST, located in Bécancour (Quebec, Canada). Insects (and other arthropods) were regularly sampled by visual observations, collection from the cadavers, and by using an entomological net and pitfall traps. Our results highlight that the decomposition process is a heterogeneous and complex process in Quebec, with cadavers showing signs of precocious desiccation/mummification. In addition, our observations confirm that the presence of superficial skin lesions accelerates the colonization of blow flies (Diptera: Calliphoridae) and, consequently, the process of decomposition. Finally, we were able to discriminate between "early colonizers" (e.g., Calliphoridae Lucilia sp. or Calliphora livida), "late colonizers" (e.g., larvae of Piophilidae or Heleomyzidae), and "non-specific colonizers." We also officially report the first observation of Cochliomyia macellaria (Diptera: Calliphoridae) in Quebec. These findings will provide new information to help medico-legal death investigations by determining the minimum time elapsed since death and the circumstances surrounding death.

Revolution in death sciences: body farms and taphonomics blooming. A review investigating the advantages, ethical and legal aspects in a Swiss context.

Taphonomy is the study of decaying organisms over time and their process of fossilization. Taphonomy, originally a branch of palaeontology and anthropology, was developed to understand the ecology of a decomposition site, how site ecology changes upon the introduction of plant or animal remains and, in turn, how site ecology affects the decomposition of these materials. In recent years, these goals were incorporated by forensic science to understand the decomposition of human cadavers, to provide a basis on which to estimate postmortem and/or postburial interval, to assist in the determination of cause and circumstances of death, and to aid in the location of clandestine graves. These goals are achieved through the study of the factors that influence cadaver decomposition (e.g. temperature, moisture, insect activity). These studies have also provided insight into the belowground ecology of cadaver breakdown and allowed to develop useful protocols for mass disaster managements in humanitarian medicine. From the results obtained, new scientific disciplines have arisen, gathered under the word "taphonomics" such as the study of microorganisms living below/on a cadaver (thanatogeomicrobiology), and join the more classical forensic sciences such as anthropology, botany or entomology. Taking into account the specificities of the study object (human cadaver), primordial requirements are needed in terms of security (physical and environmental) as well as ethical and legal concerns which are studied in the Swiss context. The present review aims to present in a first part the concept of human forensic taphonomy facilities (HFTF, also colloquially named "body farm") leading to an enrichment of forensic sciences with new "taphonomics". The second part is focused on the mandatory points that must be addressed for a HFTF approach, especially because it requires a specific place to undertake this research which must be performed in conformity with a country's human ethics and laws.

Understanding clothed buried remains: the analysis of decomposition fluids and their influence on clothing in model burial environments.

Previous studies of fabric degradation have shown promising results for post-mortem interval estimations based on differences in the degradation states of clothing in the presence of decomposing remains. It is crucial to determine if a body was present when using the degradation state as an indicator of time since death. For this study, fabric samples from buried pig remains were collected and analyzed using attenuated total reflectance Fourier transform infrared spectroscopy and chromatography- mass spectrometry. Three different fabrics were investigated; 100% cotton, 100% polyester and a polyester-cotton blend. Distinct visual changes were observed between the experimental and control graves, with the fabrics in the control grave degrading more rapidly. There was also a difference between the fabric types, whereby the natural fabrics degraded much faster than the synthetic ones. Principal component analysis was used to determine that the cotton control samples could be statistically separated based on their degradation state. The presence of lipids and proteins were useful for separating "wetter" graves from those drier in nature as well as the control graves. Clothing evidence was demonstrated to provide quantitative time since death information, as well as indicating the decomposition site in the event of intentional or unintentional movement.

Profiling the scent of weathered training aids for blood-detection dogs.

At outdoor crime scenes, cadaver-detection and blood-detection dogs may be tasked with locating blood that is days, weeks or months old. Although it is known that the odour profile of blood will change during this time, it is currently unknown how the profile changes when exposed to the environment. Such variables must be studied in order to understand when the odour profile is no longer detectable by the scent-detection dogs and other crime scene tools should be implemented. In this study, blood was deposited onto concrete and varnished wood surfaces and weathered in an outdoor environment over a three-month period. Headspace samples were collected using solid phase microextraction (SPME) and analysed using comprehensive two-dimensional gas chromatography - time-of-flight mass spectrometry (GC×GC-TOFMS). The chemical odour profiles were compared with the behavioural responses of cadaver-detection and blood-detection dogs during training. Data interpretation using principal component analysis (PCA) and hierarchical cluster analysis (HCA) established that the blood odour could no longer be detected using SPME-GC×GC-TOFMS after two months of weathering on both surfaces. Conversely, the blood-detection dogs had difficulty locating the blood samples after one month of weathering on concrete and after one week of weathering on varnished wood. The scent-detection dogs evaluated herein had not been previously exposed to environmentally weathered blood samples during training. Given that this study was conducted to test the dogs' baseline abilities, it is expected that with repeated exposure, the dogs' capabilities would likely improve. The knowledge gained from this study can assist in providing law enforcement with more accurate training aids for blood-detection dogs and can improve their efficiency when deployed to outdoor crime scenes.

The influence of ageing and surface type on the odour profile of blood-detection dog training aids.

Cadaver-detection dogs are a preferred search tool utilised by law enforcement agencies for the purposes of locating victim remains due to their efficiency and minimal disturbance to the crime scene. In Australia, a specific group of these canines are blood-detection dogs, which are trained to detect and locate blood evidence and search potential crime scenes in cases where a cadaver may not be present. Their role sometimes requires searches to be carried out after considerable time has passed since the crime occurred, and this is important for developing effective training protocols. This study aimed to investigate the volatile organic compounds (VOCs) produced from fresh and aged human blood on various surfaces. Solid phase microextraction (SPME) was used to extract VOCs from the headspace of dried blood samples aged and sampled periodically over 12 months from a non-porous (i.e. aluminium) and porous (i.e. cotton) surface. Samples were analysed using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS). Fresh blood produced distinctively different VOC patterns compared to blood aged longer than 1 week with the overall profile differing between the two surface types, and a large subset of the VOC profile found to be responsible for these differences. When analysing the various functional groups present in the samples, a common pattern between ages and surface types was observed with no specific chemical class dominating the overall profile. The results highlight the importance of evaluating training aids for scent-detection canines to ensure the greatest efficacy during training and subsequently at crime scene searches.

Inter-year repeatability study of volatile organic compounds from surface decomposition of human analogues.

Decomposition odour and volatile organic compounds (VOCs) have gained considerable attention recently due to their use by insects and scent detection canines to locate remains. However, a comprehensive and accurate profile of decomposition odour is yet to be confirmed. This is, in part, due to the geographical diversity in the studies conducted and the variation in the methodology and compounds being reported. To date, no repeatability studies of decomposition odour have been conducted in the same environment. In order to address this current gap in the scientific literature, this study conducted three replicate trials in order to evaluate the inter-year repeatability of the decomposition VOC profile in a southern Canadian environment. Surface decomposition trials were conducted during the spring and summer months and the VOCs were analysed by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). This study was able to demonstrate that decomposition VOCs are produced consistently during their characteristic stages and that this relationship is maintained under varying environmental factors which influence the rate of decomposition. This consistent production of decomposition VOCs can lead to a better understanding of the mechanisms of soft tissue decomposition and their sources of variation, and it could potentially lead to improved applications of these compounds for the detection of decomposed remains.

GC × GC-TOFMS and supervised multivariate approaches to study human cadaveric decomposition olfactive signatures.

In forensic thanato-chemistry, the understanding of the process of soft tissue decomposition is still limited. A better understanding of the decomposition process and the characterization of the associated volatile organic compounds (VOC) can help to improve the training of victim recovery (VR) canines, which are used to search for trapped victims in natural disasters or to locate corpses during criminal investigations. The complexity of matrices and the dynamic nature of this process require the use of comprehensive analytical methods for investigation. Moreover, the variability of the environment and between individuals creates additional difficulties in terms of normalization. The resolution of the complex mixture of VOCs emitted by a decaying corpse can be improved using comprehensive two-dimensional gas chromatography (GC × GC), compared to classical single-dimensional gas chromatography (1DGC). This study combines the analytical advantages of GC × GC coupled to time-of-flight mass spectrometry (TOFMS) with the data handling robustness of supervised multivariate statistics to investigate the VOC profile of human remains during early stages of decomposition. Various supervised multivariate approaches are compared to interpret the large data set. Moreover, early decomposition stages of pig carcasses (typically used as human surrogates in field studies) are also monitored to obtain a direct comparison of the two VOC profiles and estimate the robustness of this human decomposition analog model. In this research, we demonstrate that pig and human decomposition processes can be described by the same trends for the major compounds produced during the early stages of soft tissue decomposition.

Reducing variation in decomposition odour profiling using comprehensive two-dimensional gas chromatography.

Challenges in decomposition odour profiling have led to variation in the documented odour profile by different research groups worldwide. Background subtraction and use of controls are important considerations given the variation introduced by decomposition studies conducted in different geographical environments. The collection of volatile organic compounds (VOCs) from soil beneath decomposing remains is challenging due to the high levels of inherent soil VOCs, further confounded by the use of highly sensitive instrumentation. This study presents a method that provides suitable chromatographic resolution for profiling decomposition odour in soil by comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry using appropriate controls and field blanks. Logarithmic transformation and t-testing of compounds permitted the generation of a compound list of decomposition VOCs in soil. Principal component analysis demonstrated the improved discrimination between experimental and control soil, verifying the value of the data handling method. Data handling procedures have not been well documented in this field and standardisation would thereby reduce misidentification of VOCs present in the surrounding environment as decomposition byproducts. Uniformity of data handling and instrumental procedures will reduce analytical variation, increasing confidence in the future when investigating the effect of taphonomic variables on the decomposition VOC profile.

An initial investigation into the ecology of culturable aerobic postmortem bacteria.

Postmortem microorganisms are increasingly recognized for their potential to serve as physical evidence. Yet, we still understand little about the ecology of postmortem microbes, particularly those associated with the skin and larval masses. We conducted an experiment to characterize microbiological and chemical properties of decomposing swine (Sus scrofa domesticus) carcasses on the island of Oahu, Hawaii, USA, during June 2013. Bacteria were collected from the head, limb, and larval mass during the initial 145h of decomposition. We also measured the pH, temperature, and oxidation-reduction potential of larval masses in situ. Bacteria were cultured aerobically on Standard Nutrient Agar at 22°C and identified using protein or genetic signals. Carcass decomposition followed a typical sigmoidal pattern and associated bacterial communities differed by sampling location and time since death, although all communities were dominated by phyla Actinobacteria, Firmicutes, and Proteobacteria. Larval masses were reducing environments (~-200mV) of neutral pH (6.5-7.5) and high temperature (35°C-40°C). We recommend that culturable postmortem and larval mass microbiology and chemistry be investigated in more detail, as it has potential to complement culture-independent studies and serve as a rapid estimate of PMI.

Detection of decomposition volatile organic compounds in soil following removal of remains from a surface deposition site.

PURPOSE: Cadaver-detection dogs use volatile organic compounds (VOCs) to search for human remains including those deposited on or beneath soil. Soil can act as a sink for VOCs, causing loading of decomposition VOCs in the soil following soft tissue decomposition. The objective of this study was to chemically profile decomposition VOCs from surface decomposition sites after remains were removed from their primary location. METHODS: Pig carcasses were used as human analogues and were deposited on a soil surface to decompose for 3 months. The remains were then removed from each site and VOCs were collected from the soil for 7 months thereafter and analyzed by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS). RESULTS: Decomposition VOCs diminished within 6 weeks and hydrocarbons were the most persistent compound class. Decomposition VOCs could still be detected in the soil after 7 months using Principal Component Analysis. CONCLUSIONS: This study demonstrated that the decomposition VOC profile, while detectable by GC×GC-TOFMS in the soil, was considerably reduced and altered in composition upon removal of remains. Chemical reference data is provided by this study for future investigations of canine alert behavior in scenarios involving scattered or scavenged remains.

Bacterial populations associated with early-stage adipocere formation in lacustrine waters.

The preservation of soft tissue is a valuable evidence for forensic investigation as it may provide information about the cause and manner of death as well as the time since death. Adipocere forms from the conversion of triglycerides in the neutral fats into stable fatty acids producing a solid white product which aids tissue preservation. Adipocere will typically form in water-logged grave sites and aquatic environments. Documentation on the chemical and microbiological changes that cause adipocere formation in aquatic environments is scant and mostly based on observational case reports. The aim of this study was to monitor the early adipocere formation in lacustrine waters to investigate the effect of aquatic bacteria on adipocere formation. Tissue samples from pork (Sus scrofa domesticus) belly were submerged in water samples from Lake Ontario and deionised water (control). Bacteria samples from both water and tissue were harvested. Changes in the fatty acid composition of the tissue were determined using gas chromatography-mass spectrometry. Early-stage adipocere formation was confirmed on porcine tissue submerged in lake water but was not identified on porcine tissue submerged in deionised water. Adipocere formation required an abundance of gram-positive bacteria during the early postmortem period to assist in lipolysing the triglycerides into free fatty acids. Formation of adipocere in the lake water resulted in a decrease in bacterial concentrations in the tissue over time.

Effect of age and storage conditions on the volatile organic compound profile of blood.

Cadaver-detection dogs are used by the police to locate missing persons, victims of homicide, and human remains following mass disasters. Training is conducted using a variety of training aids including blood which can be hours, weeks or months old and stored under variable conditions. The aim of this study was to chemically profile human blood using solid-phase microextraction coupled with gas chromatography-mass spectrometry to determine how the volatile organic compound (VOC) profile changed over time and under variable storage conditions. The VOC profiles of fresh and degraded blood were analyzed as well as blood stored at room temperature, refrigerated, and frozen. Fresh and degraded blood samples produced distinctive VOC patterns with VOC profiles becoming more complex over time. Freezing the blood produced a complex VOC profile that was clearly discriminated from the VOC profile for blood stored at room temperature or in a refrigerator. This study highlights the importance of standardizing the age and storage conditions when using blood as a training aid to ensure cadaver-detection dogs are exposed to an accurate representation of the blood VOCs they may encounter at a scene.

The taphonomic impact of scavenger guilds in peri-urban and rural regions of central and southern Alberta. Part I - Identification of forensically relevant vertebrate scavengers.

As a body decomposes in an outdoor environment, numerous taphonomic agents can act on the process of human decomposition. It is important to understand the impact of these agents as they can vary the rate of soft and hard tissue loss which may alter postmortem interval estimations. One taphonomic factor which has not been extensively investigated in many regions of the world, including Canada, are vertebrate scavengers. The current study aimed to identify scavenger guilds in the peri-urban and rural regions of two major cities in Alberta (Calgary and Edmonton) where human remains are frequently located. Vertebrate scavenger activity was recorded continuously using cellular and noncellular trail cameras. Images were analyzed to determine how the scavenging profiles (i.e., scavenger species, arrival time, and feeding behavior) impacted the loss of soft and hard tissue. We identified a range of mammalian and avian scavengers and found that coyote and black-billed magpie were the predominant scavengers recorded at the Edmonton peri-urban and rural sites, and the Calgary peri-urban sites. In contrast, when a site was within bear territory such as the Calgary rural sites, black and grizzly bears were the predominant scavengers. At all sites, the large mammalian scavengers were responsible for most soft tissue loss and subsequent hard tissue dispersal. None of the scavengers demonstrated a clear preference for open versus closed sites. This taphonomic information is important to consider when searching for human remains at these locations or in other North American regions with comparable scavenger guilds.

Taphonomic impact of vertebrate scavengers on degradation and dispersal of remains, southeastern British Columbia.

Vertebrate scavengers represent important taphonomic agents that can act on a body, particularly when in an outdoor environment. Understanding the effects of these agents will direct how and where to search for human remains and influence the likelihood of discovery in a particular region. The current study aimed to identify the taphonomic impact of scavenger guilds in the peri-urban and rural regions of southeastern British Columbia. Vertebrate scavenger activity on pig carcasses was recorded remotely using trail cameras and analyzed to determine temporal scavenging profiles. Both the peri-urban and rural environments produced comparable scavenger guilds, namely: turkey vultures, American crows/northern ravens (classified as "corvids"), American black bears, and coyotes. Although the two locations had different study lengths due to variable degrees of scavenging, for the period that was common to both locations (summer to early fall), the black bear was the most frequent scavenger followed by coyote. However, the dispersal of remains by the mammalian scavengers was distinctly different between sites. Only 12%-33% of skeletal elements were recovered at the rural sites compared to 80%-90% recovered at the peri-urban sites, even though the latter sites had a longer study timeframe. The extended timeframe of the peri-urban sites confirmed that certain scavengers (e.g., turkey vultures and black bears) are only seasonally active in this region. These findings demonstrate the variability of scavenger behavior and the need to assign caution and local ecological knowledge when predicting scavenger trends. Such taphonomic information is relevant for human remains searches in regions with comparable scavenger guilds.

Comparison of GPR signals over simulated clandestine graves with domestic pigs (Sus Scrofa domesticus) and human remains.

Studies assessing the use of ground-penetrating radar (GPR) for locating unmarked human graves commonly use pigs as proxies, with recent concerns about the adequacy of pigs as substitutes for humans. Also, there is little agreement on how to identify and describe GPR signals associated with graves. Hence, this project's aim is to compare GPR signals acquired over simulated clandestine graves with pig and human remains. We established human, pig, and control graves at the REST[ES] human decomposition facility in May 2022 and monitored the graves over 17 months using a 250 MHz antenna GPR system. Our results showed the presence of perturbed and V-shaped reflectors, diffraction hyperbolas, and reflectors with amplitude loss at depth between 0.6 and 0.75 m in the radargram for graves with human and pig remains. We corroborate recent studies which concluded that the use of proxies is a viable alternative to human cadavers. The observed radar signatures were classified into five key patterns, which are characteristic of similar data collected with 250 MHz above graves reported in the literature. These classes are: V-shaped dipping reflections from grave walls (class A), small hyperbolic reflections superimposed onto a near-linear reflector (class B), hyperbolic reflections from remains within the grave (class C), new high-amplitude reflection patterns (class D) and significant loss or interruption of reflections (class E). Our proposed classification can help streamline future investigations where the goal is to interpret burials within large GPR datasets and provide language to communicate these results to the broader scientific community.

Characterization of volatile organic compounds from human analogue decomposition using thermal desorption coupled to comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry.

Complex processes of decomposition produce a variety of chemicals as soft tissues, and their component parts are broken down. Among others, these decomposition byproducts include volatile organic compounds (VOCs) responsible for the odor of decomposition. Human remains detection (HRD) canines utilize this odor signature to locate human remains during police investigations and recovery missions in the event of a mass disaster. Currently, it is unknown what compounds or combinations of compounds are recognized by the HRD canines. Furthermore, a comprehensive decomposition VOC profile remains elusive. This is likely due to difficulties associated with the nontarget analysis of complex samples. In this study, cadaveric VOCs were collected from the decomposition headspace of pig carcasses and were further analyzed using thermal desorption coupled to comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (TD-GC × GC-TOFMS). Along with an advanced data handling methodology, this approach allowed for enhanced characterization of these complex samples. The additional peak capacity of GC × GC, the spectral deconvolution algorithms applied to unskewed mass spectral data, and the use of a robust data mining strategy generated a characteristic profile of decomposition VOCs across the various stages of soft-tissue decomposition. The profile was comprised of numerous chemical families, particularly alcohols, carboxylic acids, aromatics, and sulfides. Characteristic compounds identified in this study, e.g., 1-butanol, 1-octen-3-ol, 2-and 3-methyl butanoic acid, hexanoic acid, octanal, indole, phenol, benzaldehyde, dimethyl disulfide, and trisulfide, are potential target compounds of decomposition odor. This approach will facilitate the comparison of complex odor profiles and produce a comprehensive VOC profile for decomposition.

Post-mortem detection of gasoline residues in lung tissue and heart blood of fire victims.

The purpose of this study was to determine whether gasoline residues could be detected post-mortem in lung tissue and heart blood of fire victims. The lungs and heart blood were investigated to determine whether they were suitable samples for collection and could be collected without contamination during an autopsy. Three sets of test subjects (pig carcasses) were investigated under two different fire scenarios. Test subjects 1 were anaesthetized following animal ethics approval, inhaled gasoline vapours for a short period and then euthanized. The carcasses were clothed and placed in a house where additional gasoline was poured onto the carcass post-mortem in one fire, but not in the other. Test subjects 2 did not inhale gasoline, were clothed and placed in the house and had gasoline poured onto them in both fires. Test subjects 3 were clothed but had no exposure to gasoline either ante- or post-mortem. Following controlled burns and suppression with water, the carcasses were collected, and their lungs and heart blood were excised at a necropsy. The headspace from the samples was analysed using thermal desorption-gas chromatography-mass spectroscopy. Gasoline was identified in the lungs and heart blood from the subjects that were exposed to gasoline vapours prior to death (test subjects 1). All other samples were negative for gasoline residues. These results suggest that it is useful to analyse for volatile ignitable liquids in lung tissue and blood as it may help to determine whether a victim was alive and inhaling gases at the time of a fire.

Toxicological Analysis of Drugs in Human Mummified Bodies and Proposed Guidelines.

From palaeopathology to forensic taphonomy, mummified human bodies constitute biological archives of paramount importance. Toxicology analysis of endobiotics and xenobiotics has already shown value to archaeological mummies research with detecting heavy metals, sedative-hypnotic drugs, and stimulants. Thanks to the large window of drug detection in hair and nails, the information from such studies has increased the scientific community's knowledge regarding past populations' lifestyles. Still, few bibliographic references exist regarding toxicology reports in mummified bodies from forensic settings. Here, the authors aim to draw attention to the valuable contribution of toxicology analysis, taking into account previously conducted studies and their findings. Given that sample collection on mummified bodies from forensic scenarios may not always happen in laboratories or autopsy rooms, the authors also suggest guidelines for in situ sampling of forensic mummies. It is expected that the present technical note will encourage experts to perform toxicology analysis in mummified bodies and publish their case reports more often.

Seasonal impact of scavenger guilds as taphonomic agents in central and northern Ontario, Canada.

The process of human decomposition is driven by biological decomposers, mainly bacteria, vertebrates, and invertebrate scavengers. When vertebrate scavengers have access to a body, they can considerably accelerate decomposition through consumption of soft tissue and dispersal of skeletal elements. Presently, there are limited data available on vertebrate scavenging activity in Canada, particularly in densely populated provinces such as Ontario. This study aimed to determine which vertebrate species belong to the scavenger guilds in central and northern Ontario, and the impact of season and habitat on these taphonomic agents. Seasonal trials were conducted in summer, fall, and spring of 2020/2021 with pig carcasses placed in open (grassland) and closed (forest) sites. Vertebrate scavenger activity was recorded continuously using cellular and non-cellular trail cameras. Photographs were analyzed to identify species, quantify feeding intensity, and document scavenging behavior. We identified four mammalian scavengers, namely coyote, red fox, fisher, and pine marten, and three avian scavengers, namely bald eagle, turkey vulture, and American crows/northern ravens (grouped as corvids) across the trials. Season impacted scavenger presence with feeding and loss of soft tissue occurring more quickly in the summer, followed by spring and fall. None of the scavengers demonstrated a clear preference for the open versus closed sites. Our findings have identified the most prevalent vertebrate scavengers in central and northern Ontario and their taphonomic impact on soft and hard tissues. It is important to consider these agents and their ability to degrade and disperse remains during the search and recovery of human remains.