DNA Barcoding Technique Research Articles

Morphological and molecular identification of freshwater eutardigrade Dactylobiotus parthenogeneticus (Bertolani, 1982) in the Greater Zab River of Kurdistan Region - Iraq.

Dactylobiotus parthenogeneticus is one of the widespread species of tardigrade all over the world. Tardigrades of this species were collected from the Greater Zab River in Erbil City-Iraq by filtering water of the river through a plankton net with a mesh of 45 µm pore. The samples were mounted on a slide with a cover slip and examined under the microscope to determine morphological characteristics and measurements. Based on these characters the species identified to be D. parthenogeneticus. To support this diagnosis, DNA barcoding techniques were applied to do molecular analysis and sequencing on the cytochrome oxidase subunit I (COI) gene. The sequence was subjected to the GenBank database of NCBI and recorded with the accession number PP140905. The result of the sequencing and molecular analysis of the cytochrome oxidase subunit I (COI) gene confirmed to be the same species diagnosed by relying upon morphological characters. This study represents one of the pioneer researches and documents on tardigrades and found D. parthenogeneticus for the first time in the Greater Zab River in Kurdistan, North of Iraq. Tardigrades play a magnificent role in different trophic levels and can be utilized as an indicator of ecosystem health.

Combating Illegal Trade of Monitor Lizards (Varanus spp) in Malaysia through DNA Barcoding Applications

Abstract Varanids, known as monitor lizards are traded as pet animal commodities and demand for skin trade especially in Southeast Asian countries. Given the significant demand in the international trade, accurate species identification is imperative for Varanids particularly endangered species to prevent misidentification and fraud in marketplaces as Varanids species exhibit morphological similarities. Therefore, advanced molecular technique is vital to facilitate accurate species identification especially for closely related species. Besides, few research have used cytochrome oxidase subunit I (COI) to track the genetic and phylogenetic analysis of Varanids. In this study, the COI mitochondrial DNA region was tested as a molecular marker for species identification of Malaysia monitor lizards using the DNA barcode. Two species were successfully identified from eight samples from Pantai Penyabong, Mersing, Johor which are V. salvator and V. nebulosus based on phylogenetic analysis (Neighbor-Joining, Maximum Parsimony and Bayesian Tree). This study showed that by using their relative genetic divergence and evolutionary links, DNA barcodes can aid in the species identification through molecular technique. Short sequences can be used to identify closely related species, and the possibility of species misidentification among the study’s samples should be taken into account. DNA barcoding techniques should be widely practised especially in wildlife forensics to facilitate precise wildlife identification and improve the efficiencies in combatting illegal wildlife trade in Malaysia.

Unravelling the Molecular Identity of Bulgarian Jumping Plant Lice of the Family Aphalaridae (Hemiptera: Psylloidea).

Psyllids (Hemiptera: Psylloidea) are plant sap-sucking insects whose identification is often difficult for non-experts. Despite the rapid development of DNA barcoding techniques and their widespread use, only a limited number of sequences of psyllids are available in the public databases, and those that are available are often misidentified. Here, we provide 80 sequences of two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (Cytb), for 25 species of Aphalaridae, mainly from Bulgaria. The DNA barcodes for 15 of these species are published for the first time. In cases where standard primers failed to amplify the target gene fragment, we designed new primers that can be used in future studies. The distance-based thresholds for the analysed species were between 0.0015 and 0.3415 for COI and 0.0771 and 0.4721 for Cytb, indicating that the Cytb gene has a higher interspecific divergence, compared to COI, and therefore allows for more accurate species identification. The species delimitation based on DNA barcodes is largely consistent with the differences resulting from morphological and host plant data, demonstrating that the use of DNA barcodes is suitable for successful identification of most aphalarid species studied. The phylogenetic reconstruction based on maximum likelihood and Bayesian inference analyses, while showing similar results at high taxonomic levels to previously published phylogenies, provides additional information on the placement of aphalarids at the species level. The following five species represent new records for Bulgaria: Agonoscena targionii, Aphalara affinis, Colposcenia aliena, Co. bidentata, and Craspedolepta malachitica. Craspedolepta conspersa is reported for the first time from the Czech Republic, while Agonoscena cisti is reported for the first time from Albania.

Peran Teknologi dalam Meningkatkan Efisiensi Pertanian

Abstracts. This research examines the role of genetic technologies in the conservation of threatened marine species with a focus on the effectiveness of methods and the successful application of advanced techniques. The background of this research is driven by the biodiversity crisis affecting marine species, which requires innovative approaches for protection and recovery. The main objective of the study was to evaluate various genetic methods in improving the conservation of threatened species and determine their success in practice. The methods used included an in-depth literature review of genetic techniques, case studies of field applications, and analysis of results obtained from various sources. Findings suggest that genetic methods, such as CRISPR and DNA barcoding techniques, have significant potential to improve the effectiveness of marine species conservation by providing solutions to specific problems related to threatened species. Implications of this research include recommendations for the application of genetic techniques in conservation programs as well as encouragement for further research to address complex conservation challenges. This research contributes to the development of more effective genetics-based conservation strategies, offering practical guidance for future policies and actions.

Geographical range extension of two penaeid shrimp Penaeus pulchricaudatus Stebbing, 1914 and Kishinouyepenaeopsis incisa (Wang & Liu in Liu & Wang, 1987) in Bangladesh waters

Shrimps are among the most intriguing groups within the Decapoda order. In Bangladesh, penaeid shrimps hold significant economic value as a major export commodity. This study aimed to analyze the morphometric and molecular characteristics of penaeid shrimps in Bangladesh. Two species of penaeid shrimp, Penaeus pulchricaudatus and Kishinouyepenaeopsis incisa were identified as new records in Bangladesh waters. A distinguishing feature of P. pulchricaudatus is its dark brown transverse bands that do not extend to the lower half of the carapace. The identification of these species was confirmed using both morphometric characteristics and DNA barcoding techniques. Two sequences were generated for these species utilizing the 16S rRNA gene marker. The findings reveal two new records of shrimp species in Bangladesh, expanding the known diversity of shrimp in the region and highlighting the presence of previously undocumented species. Dhaka Univ. J. Biol. Sci. 33(2): 111-117, 2024 (July)

Beneath the surface: DNA barcoding of shark fins in Singapore.

The global decline of shark populations, largely driven by overfishing to supply the shark fin trade, poses a significant threat to marine ecosystems. Southeast Asia, and particularly Singapore, is a key hub for the transit and trade of shark fins that contribute to the exploitation of these apex predators. Through the use of DNA barcoding techniques, this study aimed to determine what species of shark are involved in the Singapore shark fin trade. Fins were collected from markets, dried goods shops and traditional Chinese medicine halls throughout Singapore. In total, DNA was extracted from 684 fins collected in January 2024 and PCR amplification targeted a fragment of the mitochondrial COI gene for species identification. Results revealed fins from 24 species across 16 genera, with 19 species listed on CITES Appendices II, and 16 listed as threatened on the IUCN Red List (critically endangered = 2, endangered = 4, vulnerable = 10). The top five most frequently identified species were Carcharhinus falciformis, Galeorhinus galeus, Rhizoprionodon oligolinx, Sphyrna lewini and Rhizoprionodon acutus. Of these, four are listed on CITES Appendix II and four are listed as threatened on the IUCN Red List.

Identification of Euphorbiae pekinensis Radix and its counterfeit and adulterated products based on DNA barcode, UPLC-Q-TOF-MS, UPLC fingerprint, and chemometrics.

Euphorbiae pekinensis Radix (EPR) is a traditional Chinese herb commonly used to treat edema, pleural effusion, and ascites. However, counterfeit and adulterated products often appear in the market because of the homonym phenomenon, similar appearance, and artificial forgery of Chinese herbs. This study comprehensively evaluated the quality of EPR using multiple methods. The DNA barcode technique was used to identify EPR, while the UPLC-Q-TOF-MS technique was utilized to analyze the chemical composition of EPR. A total of 15 tannin and phenolic acid components were identified. Furthermore, UPLC fingerprints of EPR and its common counterfeit products were established, and unsupervised and supervised pattern recognition models were developed using these fingerprints. The backpropagation artificial neural network and counter-propagation artificial neural network models accurately identified counterfeit and adulterated products, with a counterfeit ratio of more than 25%. Finally, the contents of the chemical markers 3,3'-di-O-methyl ellagic acid-4'-O-β-D-glucopyranoside, ellagic acid, 3,3'-di-O-methyl ellagic acid-4'-O-β-d-xylopyranoside, and 3,3'-di-O-methyl ellagic acid were determined to range from 0.05% to 0.11%, 1.95% to 8.52%, 0.27% to 0.86%, and 0.10% to 0.42%, respectively. This proposed strategy offers a general procedure for identifying Chinese herbs and distinguishing between counterfeit and adulterated products.

Assessment of the status of Spodoptera species (Lepidoptera: Noctuidae: Armyworm) in India through DNA barcoding technique

Assessment of the status of Spodoptera species (Lepidoptera: Noctuidae: Armyworm) in India through DNA barcoding technique

In-silico approaches for discrimination of Curcuma species and their closely related family using the novel technique of DNA Barcoding

In this study, we have discriminated and identified the Genus of Curcuma and related species Zingiberaceae using rbcL and trnL DNA barcode primers. Curcuma genus related to the family Zingiberaceae comprises a significant number of medicinal plants renowned for their use in ethnomedicine, playing a pivotal role in the medical, health, and pharmaceutical sectors. Traditionally, morphological methods alone have proven insufficient for accurately identifying species within this family. However, DNA barcoding technology provides a contemporary solution by utilizing plant DNA sequences for species identification, thus enabling effective conservation efforts. We used DNA barcoding techniques and for analysis used the Maximum Parsimony tree in MEGA 11 with the Kimura 2-parameter (KP model) to analyse the genetic relationships between species. Out of the 13 accessions that were studied, 12 accessions belonged to Curcuma caesia and 1 accession belonged to Curcuma aeruginosa. The genetic relationships observed were correlated with the geographical distributions of these species. It was determined that C. aeruginosa is a mutated species of C. caesia. Additionally, 1 specimen of Alpinia galanga, a plant species related to the Zingiberaceae. Barcode primer trnL primer demonstrated a 92% efficiency during the investigation. The rbcL and trnL loci are recommended as potential barcode markers for discriminating between different plant species. This study developed a comprehensive DNA barcoding database that can confidently differentiate between species by combining morphological and molecular data. This database has the potential to identify adulteration in herbal products, combat illegal trade and adulteration of plant species, and assist in germplasm conservation efforts.

Combining Morphological Characteristics and DNA Barcoding Techniques Confirm Sea Urchins of the Genus Echinometra (Echinodermata: Echinoidea) in Marine Habitat Located at Extreme Regions of the Caribbean Sea.

Echinometra spp. are pantropical echinoids found in benthic marine habitat throughout the Caribbean, Atlantic, and Indo-West Pacific oceanic regions. Currently, morphology and molecular data are sparse for echinoids observed along the northeastern coast of Toco, Trinidad, where they are relatively common. Additionally, accurate species identity for Echinometra spp. remains dynamic at both northernmost and southernmost parts of the Caribbean Sea. Although distribution of sea urchins in the genus Echinometra have extensively been studied throughout the Atlantic and Indo-West Pacific, information on its range of distribution at the edge of the Caribbean Sea is lacking. In this study, the mitochondrial Cytochrome c Oxidase subunit I (mt COI) gene was amplified using polymerase chain reaction, then sequenced. Based on successfully obtained gene sequences for 581 base pairs, the echinoid species Echinometra lucunter and Echinometra viridis were identified for black and red color morphotypes from Trinidad (n=23) and Key Largo, Florida (n=6), respectively. Furthermore, these specimens were genetically identical to species identified in other studies for Puerto Rico, Panamá, Honduras, and Belize. Although morphological variations, such as spine and test color occur throughout Echinometra spp., molecular identification using the barcoding technique confirmed E. lucunter color morphs for the first time in Trinidad. Since the status of E. lucunter populations, specifically at the most northern and southern regions of the Caribbean Sea is dynamic, further studies using gene markers are essential in determining species distribution, in light of current trends in climate change.

DNA Barcoding Revealed Mislabeling of Imported Seafood Products in Thailand

Seafood mislabeling threatens customer rights and causes economic loss worldwide. The information on seafood misrepresentation in Thailand is still lacking, and the investigation and monitoring program must be well established. This study investigated the mislabeling status of imported seafood in Thailand using the DNA barcoding technique. A total of 45 imported seafood products from five distributors were included. Scientific, common, local, and market names of seafood samples were obtained from FAO and Fishbase databases. DNA was extracted, and PCR was performed using a universal primer targeting the COI gene. Species of each sample were identified with over 98% similarity based on COI sequence analysis. DNA sequence revealed 11 mislabeled samples. Among substituted species, Pangasianodon hypophthalmus and Thunnus maccoyii were found to be endangered species according to IUCN status. Products obtained from Brand-C showed the highest mislabeling rate (42.85%). The phylogenetic analysis adopted with the TIM2+F+I+G4 model showed the sequenced DNA similar to the NCBI database reference sequence. Overall, mislabeled products of imported seafood were found at the rate of 24.44%, suggesting that strict surveillance for seafood substitution should be implemented in Thailand.

Introduksi Teknologi DNA Barcoding kepada Staf UPTD Perikanan Budidaya Air Payau dan Laut Karang Intan

Partners in community service activity are UPTD Brackish and Marine Aquaculture, Karang Intan, one of the centers under South Kalimantan Province that specializes in the hatchery of local fish such as snakehead (Channa striata), common carp (Cyprinus carpio), and climbing perch (Anabas testudineus). The problem in the partner is fish farming that still relies on seeds from nature, which causes several problems, such as slow seed growth, decreased production quality, disease attacks and hereditary abnormalities that are getting higher due to interbreeding. Based on these problems, the team collaborated with the UPTD for Brackish and Marine Aquaculture, Karang Intan, to improve the genetic quality of fish by introducing DNA barcoding techniques, especially at the spawning stage. The introduction of DNA barcoding technology is carried out through (1) classical counseling and group discussions, 2) training and assistance covering technical aspects of DNA barcoding (fish DNA isolation, DNA amplification, visualization of PCR products with electrophoresis, to the sequencing). Based on the evaluation, 100% of the participants were enthusiastic and realized the importance of DNA barcoding techniques to produce superior broodstock and fish fry, especially for snakehead, common carp, and climbing perch commodities.

Genetic identification of three CITES-listed sharks using a paper-based Lab-on-a-Chip (LOC).

Threatened shark species are caught in large numbers by artisanal and commercial fisheries and traded globally. Monitoring both which shark species are caught and sold in fisheries, and the export of CITES-restricted products, are essential in reducing illegal fishing. Current methods for species identification rely on visual examination by experts or DNA barcoding techniques requiring specialist laboratory facilities and trained personnel. The need for specialist equipment and/or input from experts means many markets are currently not monitored. We have developed a paper-based Lab-on-a-Chip (LOC) to facilitate identification of three threatened and CITES-listed sharks, bigeye thresher (Alopias superciliosus), pelagic thresher (A. pelagicus) and shortfin mako shark (Isurus oxyrinchus) at market source. DNA was successfully extracted from shark meat and fin samples and combined with DNA amplification and visualisation using Loop Mediated Isothermal Amplification (LAMP) on the LOC. This resulted in the successful identification of the target species of sharks in under an hour, with a working positive and negative control. The LOC provided a simple "yes" or "no" result via a colour change from pink to yellow when one of the target species was present. The LOC serves as proof-of-concept (PoC) for field-based species identification as it does not require specialist facilities. It can be used by non-scientifically trained personnel, especially in areas where there are suspected high frequencies of mislabelling or for the identification of dried shark fins in seizures.

DNA Barcoding of Locusta migratoria manilensis (Orthoptera: Acrididae) Reveals Insights into the Species and Subspecies Differentiation

Abstract Accurate identification and classification of insect species, especially those with significant economic and ecological implications, have historically presented challenges. Migratory locusts, Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae), are notorious for their destructive impact on crops. Traditional morphological methods often face limitations in distinguishing closely related species and require taxonomic expertise. However, the emergence of DNA barcoding as a powerful tool for species identification has revolutionized the field of entomology. DNA barcoding utilizes a standardized DNA sequence, a molecular barcode, which serves as a distinct genetic signature for rapid and accurate species identification. In this study, DNA barcoding techniques were employed to identify and differentiate the migratory locust subspecies manilensis, in both its solitary and gregarious forms, as well as to determine its phylogenetic relationship with other related species within the Acrididae family. GenBank reference sequences were used to identify the locusts at the molecular subspecies level. Although the COI marker did not exhibit significant differences between the solitary and migratory forms, it was valuable in resolving the identification of L. migratoria subspecies. This lack of significant differences may be attributed to limited genetic variation of COI at the subspecies level and substantial genetic similarities between the solitary and migratory forms, likely stemming from a recent common ancestor. Nonetheless, using COI remains beneficial for subspecies identification in migratory locusts.

Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis

Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.

Artvin (Türkiye) Endemiği Campanula choruhensis Kit Tan & Sorger’in DNA Barkodlaması

DNA barcoding is the method of description of species based on gene diversity. In current studies, registration, genetic identification and protection of especially endemic plant species are carried out by DNA barcoding techniques. Molecular studies are based on the amplification and sequencing of the barcode gene region by the PCR method. Campanula choruhensis Kit Tan & Sorger is endemic and widespread in Artvin, Erzurum and around Çoruh valley passing through it. Roots of this species are used in medicinal and aromatic studies. Intense roadworks and dam constructions are carried out in and around the distribution area of this species. This situation harms the habitat of the species and comes across with its extinction. In this study, the plastid barcode gene regions (trnK-rps16 and trnL-rpl32) of the C. choruhensis was sequenced from five populations. To make the identification of this species quickly and accurately, gene sequence compared with sequences of other related Campanula L. species. In addition, gene sequences and phylogenetic relations were given.

Qualitative assessment of the flowering buds of Mesua ferrea Linn with special emphasize on HPTLC and universal DNA bar-coding technique and evaluation of its antimicrobial potential

BackgroundThe study has been scientifically exploring the powder sample and extracts of flowering buds of Mesua ferrea Linn (FBMF) based on pharmacognostic and phytochemical parameters. The medicinal plant Mesua ferrea Linn was identified by Botanical verification and the universal DNA bar-coding technique. The FBMF powder’s quality was performed by the micromeritics properties like bulk density, tapped density, angle of repose, Hausner ratio, Carr’s index, and optical microscopy method, and physicochemical evaluations were performed by the swelling index, foaming index, loss on drying, extractive values, and ash values. The extract’s preliminary phytochemical screening was accomplished by the alkaloids, flavonoids, tannins, steroids, carbohydrates, and glycoside tests. To ensure the presence of a bioactive compound of each FBMF extract by qualitative HPTLC study against the reference β-sitosterol Rf value of 0.83 was revealed at 254 nm with a developed solvent system toluene, ethyl acetate, and acetic acid (6:2:0.1) v/v. Many pharmacological benefits, including those related to wound healing, nonalcoholic fatty liver disease, analgesic, sedative effects, immunomodulatory, anticancer, antimicrobial, anti-inflammatory, anxiolytic, and atopic dermatitis, have been demonstrated by this phytosterol. An antimicrobial study was carried out by a well diffusion method and, lastly, measured minimum inhibitory concentration and compared to the marketed active pharmaceutical component. Numerous skin infections of more invasive, serious illnesses are caused by potential antimicrobial activity.ResultsThe universal DNA bar-coding of plant Mesua ferrea Linn has shown a high percentage of identity. The micrometrics properties and physicochemical evaluation of the powder sample of FBMF were in an acceptable range. The preliminary phytochemical screening shows that all extracts contain steroids and carbohydrates. Moreover, flavonoids were found in ethyl acetate and ethanol, and a qualitative HPTLC study confirmed that every extract contains β-sitosterol. N-hexane extract of FBMF shows the potential antimicrobial activity with Staphylococcus aureus, and the MIC value was observed at 0.062 mg/ml.ConclusionOur research demonstrated that the plant Mesua ferrea Linn has been successfully authenticated by a DNA bar-coding technique, and all extracts of FBMF contain β-sitosterol. The n-hexane solvent extracts have shown the potential highest antimicrobial effects compared to other extracts. These results support using n-hexane extracts as a traditional medicine for treating several diseases.Graphical

DNA Barcoding as a Plant Identification Method

In the last two decades, plant taxonomy has bloomed, following the development of a novel technique, namely, DNA barcoding. DNA barcodes are standardized sequences, ideally unique, coding or non-coding, either from the genome of the organism or from its organelles, that are used to identify/classify an organismal group; in short, the method includes amplification of the DNA barcode, sequencing and comparison with a reference database containing the relevant sequences from different species. In plants, the use a universal DNA barcode, such as COI, which is used in animals, has not been achieved so far. This review provides a comprehensive overview of the progress made in DNA barcoding within the field of plant taxonomy. It highlights the success of various barcode loci, the emergence of super barcodes from the chloroplast genome, and the overall impact of next-generation sequencing technologies on the field. The discussion of different approaches reflects the ongoing efforts to refine and optimize DNA barcoding techniques for plants, contributing to the advancement in our understanding of plant biodiversity.

DNA barcoding for the identification and authentication of medicinal deer (Cervus sp.) products in China.

Deer products from sika deer (Cervus nippon) and red deer (C. elaphus) are considered genuine and used for Traditional Chinese Medicine (TCM) materials in China. Deer has a very high economic and ornamental value, resulting in the formation of a characteristic deer industry in the prescription preparation of traditional Chinese medicine, health food, cosmetics, and other areas of development and utilization. Due to the high demand for deer products, the products are expensive and have limited production, but the legal use of deer is limited to only two species of sika deer and red deer; other wild deer are prohibited from hunting, so there are numerous cases of mixing and adulteration of counterfeit products and so on. There have been many reports that other animal (pig, cow, sheep, etc.) tissues or organs are often used for adulteration and confusion, resulting in poor efficacy of deer traditional medicine and trade fraud in deer products. To authenticate the deer products in a rapid and effective manner, the analysis used 22 deer products (antler, meat, bone, fetus, penis, tail, skin, and wool) that were in the form of blind samples. Total DNA extraction using a modified protocol successfully yielded DNA from the blind samples that was useful for PCR. Three candidate DNA barcoding loci, cox1, Cyt b, and rrn12, were evaluated for their discrimination strength through BLAST and phylogenetic clustering analyses. For the BLAST analysis, the 22 blind samples obtained 100% match identity across the three gene loci tested. It was revealed that 12 blind samples were correctly labeled for their species of origin, while three blind samples that were thought to originate from red deer were identified as C. nippon, and seven blind samples that were thought to originate from sika deer were identified as C. elaphus, Dama dama, and Rangifer tarandus. DNA barcoding analysis showed that all three gene loci were able to distinguish the two Cervus species and to identify the presence of adulterant species. The DNA barcoding technique was able to provide a useful and sensitive approach in identifying the species of origin in deer products.

DNA barcoding continues to identify endangered species of shark sold as food in a globally significant shark fin trade hub.

Shark fins are a delicacy consumed throughout Southeast Asia. The life history characteristics of sharks and the challenges associated with regulating fisheries and the fin trade make sharks particularly susceptible to overfishing. Here, we used DNA barcoding techniques to investigate the composition of the shark fin trade in Singapore, a globally significant trade hub. We collected 505 shark fin samples from 25 different local seafood and Traditional Chinese Medicine shops. From this, we identified 27 species of shark, three species are listed as Critically Endangered, four as Endangered and ten as Vulnerable by the International Union for Conservation of Nature (IUCN). Six species are listed on CITES Appendix II, meaning that trade must be controlled in order to avoid utilization incompatible with their survival. All dried fins collected in this study were sold under the generic term "shark fin"; this vague labelling prevents accurate monitoring of the species involved in the trade, the effective implementation of policy and conservation strategy, and could unwittingly expose consumers to unsafe concentrations of toxic metals. The top five most frequently encountered species in this study are Rhizoprionodon acutus, Carcharhinus falciformis, Galeorhinus galeus, Sphyrna lewini and Sphyrna zygaena. Accurate labelling that indicates the species of shark that a fin came from, along with details of where it was caught, allows consumers to make an informed choice on the products they are consuming. Doing this could facilitate the avoidance of species that are endangered, and similarly the consumer can choose not to purchase species that are documented to contain elevated concentrations of toxic metals.