Wildlife forensic science is forensic science applied to legal issues involving wildlife. Wildlife forensic sciences also deal with conservation and identification of rare species and is a useful tool for non-invasive studies. Methods can be used to determine relatedness of the animals in the area allowing them to determine rare and endangered species that are candidates for genetic rescue. Techniques using things such as the SSCP or Single-Strand Conformational Polymorphism gel electrophoresis technique, microscopy, DNA barcoding, Mitochondrial Microsatellite Analysis and some DNA and Isotope analysis can identify species and individual animals in most cases if they have already been captured . Unlike human identification, animal identification requires determination of its family, genus, and species, and sex in order to individualize the animal, typically through the use of DNA based analyses.

History

Wildlife forensic science stems from the various issues that are dealt with when it comes to wildlife crime. Wildlife crime includes actions such as wildlife trafficking, poaching, wildlife cruelty, and habitat destruction.^[1] Wildlife Crime can basically be anything that threatens the existence of a species; animals, plants, bacteria, or fungi.^[2] Wildlife Crime also creates a variety of problems for wildlife. Including but not limited to ecological stability, economies, public health, and criminal justice.^[2] Out of international crime, Wildlife Crime is the third largest illegal trade behind drugs and firearms, and potentially makes $20 billion dollars per year.^[3][4] The two main ways to prevent wildlife crime is various types of legislation which helps protect and identify species, and the application of wildlife forensic science, which is used to view biological aspects of the crime, and is used as supporting evidence to pass various legislative acts.^[2] Wildlife forensic science have helped support the creation of various wildlife protection acts, such as the Endangered Species Act and the Lacey Act among others.^{[citation needed]}

Endangered Species Act

The Endangered Species Act or ESA developed in the United States. United States President, Richard M. Nixon signed the Endangered Species Act on December 28, 1973.^[5] The overarching goal of the ESA was to conserve and protected both wildlife as well as protecting their habitats across the globe. This act aimed to prevent extinction and encourage recovery of organisms, and also included protection for various ecosystems that wildlife resides in. This act developed from wildlife being threatened in the United States, and also encouraged the creation of various treaties with countries across the globe in order to protect species. This included migratory bird treaties with Canada, Mexico, and Japan and the creation of a convention for Natural Protection and Wildlife Preservation.^[6]

Lacey Act

The Lacey Act was developed in 1900, and has since gone through various expansions throughout the years to protect and enforce wildlife laws. The act initially stated with preventing hunters from killing game illegally, and then transferring them across states. The Lacey act has went through various amendments, including in 2008 and 2009, which expanded the acts reaches. These new updates included expanding to protection timber and timber products. Now, the Lacey act focuses on the illegal trafficking of not just wildlife, but plants as well. Importing, exporting, transportation, and the sale and purchase of species are now all sections that are included in the modern Lacy Act.^[7]

Migratory Bird Treaty Act

The Migratory Bird Treaty Act was enacted in 1918, and worked to implement treaties from Canada (1916), Mexico (1936), Japan (1972), and Russia (1976) to protect migratory bird populations.^[8] Migratory birds are protected if it meets any one of the following three criteria.

1. It occurs in the United States naturally, or is listed as one of the birds on the international treaties with Canada, Mexico, Japan, and Russia.^[9]
2. If there are taxonomy splits that result in a new species coming from a species that was previously on the list, and fits the criteria for number 1, they will be protected.^[10]
3. If there is new evidence that the bird used to have a natural occurrence in the United States.

Marine Mammal Protection Act

The Marine Mammal Protection Act protects a variety of mammals that use the ocean as a main survival resources.^[11] This can include mammals that live completely in the ocean such as whales, manatees, and dolphins, and animals that spend rely on the ocean, but also stay on land at times, such as walruses and polar bears.^[11] The MMPA protects against any form of collection or killing in U.S waters or by U.S citizens.^[11]

Threats

Wildlife trafficking

Wildlife trafficking includes the trading of live animals or parts of wildlife that are sold and came from the wild. Wildlife trade generates a large amount of revenue each year, and can equal billions of dollars. Various animals are trafficked for the pet trade, such as birds, reptiles, and corals. Animal parts that are commonly traded include bushmeat, animal horns for medicinal and ornamental purposes, and products to make fashion products. A prime example of a trafficked animal are pangolins, that are often trafficked for their scales. Animals across all taxa are affected by wildlife trafficking. Wildlife forensic science comes into play throughout wildlife trafficking as scientists use DNA to try and determine information about the species that are being trafficked.^{[citation needed]}

Poaching

Poaching is a complex and global problem. Part of what makes poaching such a complex issue it provides complications to conservation, additionally, poaching is also linked to poverty. However, thousands of species are faced with poaching, including animals like African Elephants and Rhinoceros.^[12] Products from poaching can include ivory, animal skins, bones, bushmeat. These items may be sold as they are or turned into leathers, traditional medicines, ornaments, and more. In addition, poaching isn't just a wildlife problem, poaching also occurs for ornamental plants as well.^{[citation needed]}

Animal cruelty

After the 2020 production of Tiger King: Murder, Mayhem, and Madnese, people are more aware of roadside zoos. These zoos have brought to light possible flaws in legislation and the limited protection of endangered wildlife. However beyond the legislative implications of operations such as roadside zoos, various wildlife crime aspects lead to animal cruelty and animal abuse.^[13] These situations often lead to animals being killed, which lends itself to wildlife forensic sciences, to explore the aftermath of these events. Wildlife forensics can assist in determining what species of animals may have been in a location, as well as determining what may have happened to wildlife if they are killed by cruelty. This can lead to convictions in cases against people operating roadside zoos and general wildlife cruelty. In addition, investigations of animal cruelty can lead to new legislation to protect wildlife.^{[citation needed]}

Habitat destruction

Habitat destruction is an additional threat that faces wildlife. Habitat destruction includes habitat fragmentation, introduction of invasive species, and true habitat destruction. However, in order to help combat habitat destruction genetic sequencing and classification of morphological structure play key roles in protecting an area.^[14] Wildlife forensic sciences have been used to sequence animals such as pangolins, and plants such as orchids, in order to identify the species living in areas that are being destroyed, and to help provide evidence and the support for protection of areas. Naming species is a key issue in being able to conserve an area, and wildlife forensics can assist in this via genetic analysis.^{[citation needed]}

Types of evidence

In order to understand wildlife crime, and in order to complete wildlife forensic science, evidence is needed. There are various parts of evidence that are used in order to understand the crime and species being affected. Evidence can take a variety of forms such as the entire organism, both living or dead, pieces of the animals (fur, feathers, bones, and organs), or even the products created from the organism. These products may include jewelries, processed meats, clothing, ornaments, and medicines.^{[15][citation needed]}

Techniques

Single-Strand Conformational Polymorphism Gel electrophoresis

A simple and sensitive technique used to identify any mutations and also used in the genotyping of animals. The technique uses the method based on the fact that single-stranded DNA has a defined conformation. Any altered conformation due to a single base change in the sequence can cause single-stranded DNA to migrate differently under nondenaturing electrophoresis conditions, so a wild-type and mutant DNA samples display different band patterns.^[16]

There are 4 steps to this method:

1. polymerase chain reaction (PCR) amplification of DNA sequence of interest
2. denaturation of double-stranded PCR products
3. cooling of the denatured DNA (single-stranded) to maximize self-annealing
4. detection of mobility difference of the single-stranded DNAs by electrophoresis under non-denaturing conditions.

DNA and isotope analysis

DNA analysis is used to help determine the species of an animal they use DNA nucleotide sequencing as a key method and follow it up by comparing sequenced DNA fragments with reference DNA sequences of different species. The similarity or sequence homology between the unknown and reference sequences facilitates to ascertain the species of origin.^[17] This technique is used to determine relatedness of a rare species and to also check for any signs of inbreeding depression in the target species to see if it is a candidate for genetic rescue. Isotope analysis is used in this same vein to determine the composition of the habitat that animal resides in.^{[citation needed]}

Mitochondrial microsatellite analysis

Mitochondrial microsatellite analysis methods are often performed to individualize^[18] the remains of an animal and determine if a species is endangered,^[19] or if it was hunted out of season.^[20] Mitochondrial DNA reference profiles can be easily be obtained from public databases like the International Nucleotide Sequence Database (INSDC), the European Molecular Biology Laboratory (EMBL), and the Bardode of Life Data System (BOLD or BOLDSystems).^[21] Mitochondrial DNA is used due to its high copy number, and the presence of differences in mutation rates among closely related species.^[18] The cytochrome c oxidase unit 1 (CO1) region (also known as the DNA barcode region^[22]) mutates at a lower rate and is used for higher level taxonomic classifications^[18] whereas the control region and cytochrome b are used in distinguishing closer related taxa due to their mutation rate being higher.^[18]

DNA barcoding

DNA barcoding is often used in Wildlife Forensic Science cases to identify an unknown species found at a crime scene.^[23] Blood, hair, bone, and other genetic materials are first collected at the scene, then DNA extraction is performed on the samples collected.^[24] After that, DNA quantification or PCR is performed to quantify the DNA, then DNA sequencing is performed to sequence the DNA.^[24] Lastly, the sequenced DNA is compared to a DNA database for a possible identification of the unknown species.^[24] This technique is often used in poaching cases, animal abuse cases, and killing of endangered animals.^[25][26]

Microscopy

This technique is when genetic microscopes are used to look down to a single cell it is used to look at recombination also look for mutations in genes it has been used to help identify many deleterious alleles in genes.^[27]

Ballistics

The science of wound ballistics is beginning to gain attention for wildlife forensics as a method to determine what type of firearm may have killed an animal.^[28] This focuses on specifically wound ballistics, and what the wound damage is on the body of the organism.^[28] These can be traced back to specific types of bullets and firearms, and may be useful in tracing crime back to certain parties or organizations.^[28]

Fingerprinting

Fingerprinting is a current technique that is actually particularly common in human crime, and overtime has begun to migrate to the wildlife forensics world.^[29] Fingerprinting can pick up a variety of marks, and beyond just fingerprints can pick up impressions of most body parts.^[29] These fingermarks can be found on most surfaces, and can either be patent or latent fingerprints. Patent fingerprints can be collected by photography, as patent fingerprints are visible to the naked eye.^[29] For latent prints, there are various methods to collect them, including powders, fuming, chemical, optical, and instrumental methods.^[29] In wildlife forensic science, fingerprinting has been used to lift latent marks off of pangolin scales, and additionally studies have recovered fingermarks on raptor feathers using magnetic and fluorescent powders.^[29] New successes and studies are also being found pulling fingerprints off of eggshells, ivory, teeth, bone, leathers, and antlers.^[29]

Forensic entomology

Forensic entomology is the use of insects to demine information about criminal cases. Forensic Entomology is commonly accepted in legal cases and is particularly helpful in determining time of death for both human and wildlife crimes.^[2] One of the key insects in these studies are blow flies, which deposit eggs on bodies, and the time off hatching for the eggs can be important for determining time of death.^[2] An example of forensic entomology occurred in Manitoba, Canada where two young black bears were found disemboweled.^[30] Officers collected blow fly eggs from the deceased cubs, and data was used to determine time of death. This data was used in the conviction and proved itself valuable in a wildlife crime context.^[30]

Forensic pathology

Forensic pathology developed out of the veterinary profession, and begin as a way to study disease in domestic animals, and eventually migrated to wildlife.^[31] In a forensic sense wildlife pathology has been used to look into the cause and the circumstances of deaths of threatened species.^[31] This forensic pathology helps provide baseline data and basic samples such as blood of feces. Forensic Pathology also includes full biopsies which can help analyze tissue and organ changes that may have lead to the death of an animal.^[31] One of the most common examinations for Forensic Pathology are necropsies, but there are also pre-mortem examinations occur as well.^{[citation needed]}

Laboratories and organizations

Various laboratories and organizations have formed in order to develop and perform wildlife forensic sciences. The Society for Wildlife Forensic Science and Scientific Working Group for Wildlife Forensic Sciences were both formed in 2011. In addition, the Wildlife Forensic and Conservation Genetics Cell was formed by merging a Wildlife Forensic and Conservation Genetics Labs. The Wildlife Forensic and Conservation Genetics Cell was formed in order to support the enforcement and creation of the Wildlife Protection Act. In addition to various laboratories, there are several organizations that also aid in the wildlife forensic science. The World Wildlife Fund helps provide education about wildlife crime and wildlife forensic sciences. CITES, TRAFFIC, and the IUCN also support wildlife forensic science, and use the data from it to support the conservation of wildlife. Finally, Interpol, an organization that handles international crime focuses specific support to wildlife forensic science and its use in solving wildlife crimes.^[32]

Scope

While animals and plants are the victims in the crimes of illegal wildlife trade and animal abuse, society is also affected when those crimes are used to fund illegal drugs, weapons and terrorism. Links between human trafficking, public corruption and illegal fishing have also been reported. The continued development and integration of wildlife forensic science as a field will be critical for successful management of the many significant social and conservation issues related to the illegal wildlife trade and wildlife law enforcement.^{[citation needed]}