Species identification from dried snake venom

Illegal trade in snake parts has increased enormously. In spite of strict protection under wildlife act, a large number of snakes are being killed ruthlessly in India for venom and skin. Here, an interesting case involving confiscation of crystallized dried snake venom and subsequent DNA-based species identification is reported. The analysis using the universal primers for cytochrome b region of the mitochondrial DNA revealed that the venom was extracted from an Indian cobra (Naja naja). On the basis of this report, the forwarding authority booked a case in the court of law against the accused for illegal hunting of an endangered venomous snake and smuggling of snake venom. This approach thus has immense potential for rapid identification of snake species facing endangerment because of illegal trade. This is also the first report of DNA isolation from dried snake venom for species identification.     

The Nature and Extent of Conservation Policing: Law Enforcement Generalists or Conservation Specialists?

Little is known about law enforcement agencies with responsibility for fish, wildlife, and natural resource protection. Traditional conceptions of fish and wildlife police associate these officers almost entirely with fishing and hunting activities. The relative scarcity of academic literature regarding fish and wildlife police agencies results in a lack of understanding about their role in the criminal justice system in general. The current study seeks to contribute to the limited existing literature by examining conservation policing in Florida, a state not yet studied in this context, by analyzing some 2,910 field events. Findings indicate that officers devote a considerable portion of their attention to traditional law enforcement activities in addition to the type of enforcement customarily associated with fish and wildlife law enforcement.     

Validation of a multiplex PCR assay for the forensic identification of Indian crocodiles

A dependable and efficient wildlife species identification system is essential for swift dispensation of the justice linking wildlife crimes. Development of molecular techniques is befitting the need of the time. The forensic laboratories often receive highly ill-treated samples for identification purposes, and thus, validation of any novel methodology is necessary for forensic usage. We validate a novel multiplex polymerase chain reaction assay, developed at this laboratory for the forensic identification of three Indian crocodiles, Crocodylus palustris, Crocodylus porosus, and Gavialis gangeticus, following the guidelines of Scientific Working Group on DNA Analysis Methods. The multiplex PCR was tested for its specificity, reproducibility, sensitivity, and stability. This study also includes the samples treated with various chemical substances and exposed to various environmental regimes. The result of this validation study promises this technique to be an efficient identification tool for Indian crocodiles and therefore is recommended for forensic purposes.     

Molecular Identification of Three Indian Snake Species Using Simple PCR–RFLP Method*

Abstract: Three endangered Indian snake species, Python molurus, Naja naja, and Xenochrophis piscator are known to be significantly involved in illegal trade. Effective authentication of species is required to curb this illegal trade. In the absence of morphological features, molecular identification techniques hold promise to address the issue of species identification. We present an effective PCR–restriction fragment length polymorphism method for easy identification of the three endangered snake species, Python molurus, Naja naja, and Xenochrophis piscator. A 431‐bp amplicon from cytochrome b gene was amplified using novel snake‐specific primers following restriction digestion with enzymes Mbo II and Fok I. The species‐specific reference fragment patterns were obtained for the target species, which enabled successful identification of even highly degraded shed skin sample confirming the utility of the technique in case of poor‐quality DNA. The assay could be effectively used for forensic authentication of three Indian snake species and would help strengthen conservation efforts.     

Profiling pythons to combat common illegal wildlife activities

Reptiles are the most common illegally traded vertebrates leading to many species becoming extinct or highly endangered; yet these species receive relatively low publicity and few genetic tests are available. Sought after as unusual pets and trophies, reptilian skin and meat are also valuable commodities, while certain anatomical features are valued in traditional medicinal cultures. The Australian carpet python is a popular target for illegal trade, breeding and export activities. It provides a model example for which to develop forensic markers that will positively impact wildlife criminal investigation and enforcement both in Australia and worldwide. We report on the initial isolation and characterisation of 24 polymorphic Short Tandem Repeat (STR) loci to enable individualisation and paternity testing of carpet pythons. These novel loci have been examined for their polymorphic content, heterozygosity and species specificity. The loci are being arranged into multiplex reactions exhibiting heterozygote balance with subsequent determination of stutter measurements. Allelic frequency databases of native populations are being developed that will allow not only match probabilities but also will potentially predict geographic origin of poached individuals. This profiling system will be subject to rigorous validation for use in legal proceedings.     

DNA检验技术在野生动物保护中的应用

随着生物技术的发展,法医DNA检验技术在保护野生动物的工作中得到了越来越多的应用.本文介绍了全球野生动物犯罪现状及保护措施,简述了法医DNA检验技术在野生动物物种鉴定、来源地确认、个体识别和亲子鉴定等方面的实际应用,重点评述了法医DNA技术用于野生动物保护的几种常用检验方法以及各自的优缺点,探讨了法医DNA技术用于野生...     

An overview of seized illegal wildlife entering the United States

The current study analyses seizures made at US ports of entry between 2003 and 2013, with the aim to identify concentrations of illegal wildlife imports into the United States. Findings show that 94% of species seized belong to six groups – mammals, molluscs, birds, reptiles, fish and coral – with mammals and reptiles making up more than half of all seizure incidents. Additionally, most seized wildlife is imported as leather products, medicinal products and as meat. The majority of seizures emanate from six countries, and illegal wildlife is primarily brought to the US via airline baggage. Temporal trends of wildlife seizures point to increases in the seizures of all groups of species, with the exception of birds. Based on these findings, we recommend using situational crime prevention techniques at US ports of entry to reduce opportunities that enable this trade.     

Species identification of Kachuga tecta using the cytochrome b gene

A DNA technique has been established for the identification to species level of tortoises. The test on the shell of the animal was used to identify samples from the species Kachuga tecta. A total of 100 tortoise shell specimens collected from the National Council of Agriculture (COA), Taiwan, were used in this study. Primer pairs were designed to amplify partial DNA fragments of cytochrome b within the mitochondrial genome. The DNA data showed that among the 100 samples, there were four distinct haplotype DNA sequences, within which there were a total of 90 variable sites. Between haplotypes I and II, there was only 1 nucleotide difference at position 228. Between haplotypes I and III, 65 nucleotide differences were observed; haplotypes I and IV, 62 nucleotide differences; and haplotypes III and IV, 56 nucleotide differences were observed. There were 66 and 63 nucleotide differences between haplotypes II and III and haplotypes II and IV respectively. All four haplotypes were compared with the DNA sequences held at the GenBank and EMBL databases. The most similar species were K. tecta (haplotype I and II), Morenia ocellata (haplotype III) and Geoclemys hamiltonii (haplotype IV), and their respective mtDNA similarities were 99.5%, 99.3%, 89.9% and 99.5%. However, as haplotype III was only 89.9% homologous with M. ocellata, it would seem that this haplotype shows only a limited relationship with a similar species registered currently in these databases. The method established by this study is an additional method for the identification of samples protected under Convention International Trade in Endangered Species (CITES) and will improve the work for the preservation of the endangered species.     

线粒体DNA cytb和HVI的分析用于种属鉴定

目的以线粒体DNA为目标序列,探讨生物检材的种属来源问题。方法复合扩增线粒体DNA细胞色素b基因(Cb)片段和D-环HVI上人源特异性DNA片段,2%琼脂糖凝胶电泳检测复合扩增产物谱带;用常规测序技术获得种属来源不明的检材Cytb基因序列,登陆美国国家生物信息中心网站主页(http://www.ncbi.nlm.nih.gov),将Cytb序列的测序结果用BLAST2.2.9[2004.5.1]进行匹配查询,查询数据库中存在的与其相匹配的物种条目。结果检材经复合扩增后电泳检测可区分人源性检材和非人源性检材;用生物信息法可确定检材种属来源结论检测线粒体DNA细胞色素b基因和D-环HVI的有关序列可在DNA分子水平上鉴别人源性检材和非人源性检材,结合测序的分子生物信息学方法,可对检材进行种属鉴定。     

Resurrecting the Dodo: Positive control DNA for species identification

Positive controls are necessary standards for inclusion in forensic tests. When working as expected they demonstrate that methods have been applied correctly, and therefore results can be interpreted with confidence. However, the requirement for positive controls can also introduce problems. For species identification in wildlife DNA forensic testing, it is possible that the DNA sequence of the case sample will be a 100% match to the positive control. Whilst clear results for negative controls will indicate that cross-contamination is unlikely, it would be preferable to have a positive control that will not appear in casework. In addition, for many endangered species, obtaining positive control DNA for species-specific testing can be problematic. Here we present a simple method to use artificially generated positive control DNA from the extinct Dodo, Raphus cucullatus, for four species-identification tests run routinely in UK wildlife forensic casework.     

Application of species-specific polymerase chain reaction in the forensic identification of tiger species

Globally, tigers are considered to be endangered, and are listed on Appendix I of CITES. A simple test, using a species-specific primer pair, was developed to identify tiger meat, faeces and dried skin, and provide forensic evidence of illegal wildlife trade. The specific fragment of mitochondrial cytochrome b gene was also successfully amplified from raw DNA products extracted from single tiger hairs. This PCR-based approach opens a new avenue to forensic identification of less-than-optimal samples.     

Phylogenetic Evidence for a Case of Misleading Rather than Mislabeling in Caviar in the United Kingdom

Sturgeons and paddlefish are freshwater fish which are highly valued for their caviar. Despite the fact that every single species of sturgeon and paddlefish is listed under CITES, there are reports of illegal trade in caviar where products are deliberately mislabeled. Three samples of caviar purchased in the United Kingdom were investigated for accurate CITES labeling using COI and cyt b sequencing. Initial species identification was carried out using BLAST followed by phylogenetic analyses using both maximum parsimony and maximum likelihood methods. Results showed no evidence for mislabeling with respect to CITES labels in any of the three samples, but we observed clear evidence for a case of misleading the customer in one sample.     

Wildlife Crime: The Problems of Enforcement

Environmental and wildlife crime appear recently to be benefitting from an increasing profile amongst those agencies tasked with their control, as well as receiving growing criminological attention. Despite this, those with responsibilities in this area report that it remains marginalised, receiving limited resources and suffering from a lack of political impetus to push such problems higher up the agenda. This is particularly so for those agencies, such as the police, that may be seen to have many more pressing objectives. This discussion paper considers the problems of relying on an enforcement approach to controlling such offences, taking, as an example, those activities that may be termed ‘wildlife crime’, focusing on the situation in England and Wales. Firstly, the legislative framework that criminalises harm or exploitation of wildlife is presented, alongside the main enforcement methods used. Next, the problems facing an enforcement approach are critically considered, the key issues being: under-resourcing and marginalisation, the large ‘dark-figure’ of wildlife crime, the possibility of corruption, the lack of seriousness with which such crimes are viewed, and the lack of deterrent effect. Finally, responses to the problems of enforcement are presented, categorised as either methods to improve enforcement or, as the author advocates, methods which are alternatives to enforcement (such as adopting a crime prevention approach). The paper concludes with suggestions for future research in this field.     

Progress of DNA-based Methods for Species Identification

Species identification of biological samples is widely used in such fields as forensic science and food industry. A variety of accurate and reliable methods have been developed in recent years. The cur...     

Age estimation charts for a modern Australian population

The ivory industry is the single most serious threat to global elephant populations. A highly sensitive, species-specific real-time PCR assay has been developed to detect and quantify African elephant (Loxodonta africana), Asian elephant (Elephas maximus) and Woolly Mammoth (Mammuthus primigenius) mitochondrial DNA from highly processed samples involved in the international ivory trade. This assay is especially useful for highly processed samples where there are no distinguishing morphological features to identify the species of origin. Using species-specific Taqman(?) probes targeting a region of the mitochondrial cytochrome b gene, we developed an assay that can be used to positively identify samples containing elephant or Woolly mammoth DNA faster and more cost-effectively than traditional sequencing methods. Furthermore, this assay provides a diagnostic result based on probe hybridization that eliminates ambiguities associated with traditional DNA sequence protocols involving low template DNA. The real-time method is highly sensitive, producing accurate and reproducible results in samples with as few as 100 copies of template DNA. This protocol can be applied to the enforcement of the Convention on the International Trade of Endangered Species (CITES), when positive identification of species from illegally traded products is required by conservation officers in wildlife forensic cases.     

“Command and Control”: 75 Years of Quasi Wildlife Policy Analysis of Ethiopia

Ethiopia’s wildlife history is at loggerheads with the theory of “environmental colonialism.” Ethiopia sets a center stage for the debate of what could have been the fate of African wildlife if Africa had not been colonized. In the 1960s, Ethiopia had a huge wildlife diversity. As of present, its wildlife is severely deteriorated, while that of the colonized countries are in much better condition. The disparity of conservation between the colonized countries and independent Ethiopia prompted this study. In the past 77 years, several wild species in Ethiopia have gone extinct, while some moved to threatened categories and their habitat vanished or deteriorated. Overall, Ethiopia currently hosts six critically endangered, 23 endangered, and 70 vulnerable species of wild animals. The numbers do not include plants, fungi, microorganisms, or invertebrates. This shows a clear pathology of the command and control policy of the wildlife conservation system of Ethiopia. The objectives of this article were to analyze previous legal documents published since 1944, up to the recent wildlife proclamation; to bring out strengths and weaknesses of these policies; and to propose possible alternative adaptive management strategies based on other similar studies. Policy instruments since 1944 were collected. This include the Imperial regime (1941–1974), the Military Government of Socialist Ethiopia (Derg) Regime (1974–1991), and the Federal Democratic Republic of Ethiopia (1991–2018). Documents were collected from two sources: the Internet and government printing press. Policy analysis involved a documentary analysis method on how Ethiopian governments’ policies addressed wildlife conservation of Ethiopia since 1944. To achieve the overall research objective, questions were formulated and guided the research direction. The analysis showed that there are good aspects of colonial wildlife conservation policies to learn from. Independence was not a guarantee for wildlife conservation. Existing national parks of East Africa including Ethiopia are the result of UNESCO missions to Africa.     

Canada and the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES): Lessons Learned on Implementation and Compliance

Liverpool Law Review - Unsustainable and illegal wildlife trade are contributing to the unprecedented levels of biodiversity loss and possible extinction of one million species. Law enforcement and...     

Privatising the green police: the role of NGOs in wildlife law enforcement

This article examines the role of NGOs in wildlife law enforcement, drawing on empirical research conducted within UK environmental law enforcement but also drawing on a review of the academic literature and policy debates concerning NGOs and pressure group activity. It examines the theoretical basis for NGO actions and different policy perspectives, the ideologies employed by NGO's and how these manifest themselves in law enforcement policies and practice. While the focus of this essay is UK and US NGO activity with its professional enforcement activity and policy networks, the impact of wildlife and environmental NGOs is significant in a range of jurisdictions and is a vital component in effective policing of wildlife legislation.     

扩增TP53内含子8用于生物检材的种属鉴定

目的扩增常见动物TP53内含子8片段,确定其在法医学生物检材种属鉴定中的应用价值。方法收集标本为包括人在内的15种常见动物的血痕或肌肉组织,提取DNA后定量,应用PCR扩增TP53内含子8,PAGE电泳,银染后观察结果。结果人和猕猴都扩增出一条长度为460bp的片段,鳝鱼、鲢鱼、青蛙、鸭、兔、猫、小白鼠、豚鼠、猪、牛、羊虽有扩增产物,但不在分型区内,鸡、狗未见扩增产物。结论扩增TP53内含子8进行种属鉴定,方法简单,灵敏度较高。     

A Review of “Taking Back Eden: Eight Environmental Cases That Changed the World”

Abstract

The biological aspects of illegal harvests of threatened wildlife are outlined. It is shown that local agriculturalists are beneficiaries of illegal harvesting and that competition from agriculture exacerbates the extinction risk. Illegal harvesting of wildlife is driven by the profitability of the exercise, but law enforcement activity can deter poaching by reducing the associated expected profits. Law enforcement may be unable to limit illegal harvesting to levels threatened populations can sustain as a result of perverse consequences or strategic responses by poachers to law enforcement activity. Poaching activity is sensitive to the beliefs of participants about future prices and the availability of wildlife. Erroneous beliefs result in price collapses being observed. Integrating legal markets with increased local control of wildlife and punitive law enforcement strategies may be the most effective and efficient means to constrain illegal harvests.