The Species and Origin of Shark Fins in Taiwan’s Fishing Ports,Markets, and Customs Detention: A DNA Barcoding Analysis

The increasing consumption of shark products, along with the shark’s fishing vulnerabilities, has led to the decrease in certain shark populations. In this study we used a DNA barcoding method to identify the species of shark landings at fishing ports, shark fin products in retail stores, and shark fins detained by Taiwan customs. In total we identified 23, 24, and 14 species from 231 fishing landings, 316 fin products, and 113 detained shark fins, respectively. All the three sample sources were dominated by Prionace glauca, which accounted for more than 30% of the collected samples. Over 60% of the species identified in the fin products also appeared in the port landings, suggesting the domestic-dominance of shark fin products in Taiwan. However, international trade also contributes a certain proportion of the fin product markets, as four species identified from the shark fin products are not found in Taiwan’s waters, and some domestic-available species were also found in the customs-detained sample. In addition to the species identification, we also found geographical differentiation in the cox1 gene of the common thresher sharks (Alopias vulpinus), the pelagic thresher shark (A. pelagicus), the smooth hammerhead shark (Sphyrna zygaena), and the scalloped hammerhead shark (S. lewini). This result might allow fishing authorities to more effectively trace the origins as well as enforce the management and conservation of these sharks.     

Performance of iSharkFin in the identification of wet dorsal fins from priority shark species

The past decade has seen a considerable rise in international concern regarding the conservation status of sharks and rays. The demand for highly prized shark commodities continues to fuel the international trade and gives fisheries incentive to use these resources, which have a low intrinsic capability to recover. Recognising the urgency for regulation, many countries voted to include more shark and ray species in the Appendices of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). However, the identification of fins in fisheries landings before they enter international trade is a major limitation for CITES compliance. This study reports the current performance of the iSharkFin system, a machine learning technology which aims to allow users to identify the species of a wet shark dorsal fin from its image. Photographs of 1147 wet dorsal fins from 39 shark species, collected in 12 countries, were used to train the algorithm over a four-year period. As new cohorts of images were used to test the performance of the learning algorithm, the accuracy of species assignments of known specimens was variable but did increase, reaching 85.3% and 59.1% at genus and species level respectively. The accuracy in predicting CITES-listed sharks versus unlisted sharks was 94.0% based on the 39 species currently represented in the baseline. Our results suggest that if supplied with high data inputs for specific fisheries assemblages and accompanied by user training, iSharkFin has promise for site-specific development as a rapid field identification tool in fisheries monitoring, and as a screening tool alongside traditional field morphology to detect potential CITES specimens for fisheries compliance and enforcement.     

Characterization of the pelagic shark‐fin trade in north‐central Chile by genetic identification and trader surveys

Shark fins have become a highly valued commodity with the major Asian fin‐trade centres supplied from global sources, including Chile. With growing concerns about the resilience of shark populations to heavy fishing pressure, there is a need for better information on shark landings to aid management efforts. In the widespread absence of shark landing records especially by species, monitoring the fin trade has been proposed as a way to assess species exploitation levels. Here, the first species assessment of the Chilean shark‐fin trade was provided. The goals of this study were to (1) determine the species composition and relative species proportion of sharks utilized in the fin trade, (2) determine the relationship between fin trader market names and species and (3) assess trader accuracy in identifying shark fin species based on fin photographs. Fins were analysed from two different fin drying facilities (n = 654) (secaderos) and two fin‐storage warehouses (n = 251). In contrast to official government landing records that only document four species in the landings, molecular species identification of the fins demonstrated that at least 10 pelagic shark species are present in the north‐central Chilean shark fin trade: Alopias superciliosus, Alopias vulpinus, Carcharhinus obscurus, Galeorhinus galeus, Isurus oxyrinchus, Isurus paucus, Lamna nasus, Prionace glauca, Sphyrna lewini, Sphyrna zygaena. The species composition of the fins from the secaderos was P. glauca (83·9%), I. oxyrinchus (13·6%), L. nasus (1·7%) and A. superciliosus (0·2%). There was generally good agreement between market names and single shark species for the trade categories ‘Azulejo’, ‘Tiburon’, ‘Tintorera’, ‘Cola de zorro’ and ‘Martillo’. In contrast, the market category ‘Carcharhinus’ consisted of a mixture of at least five species. The molecular results also identified two species (S. lewini and I. paucus) not previously recorded in Chilean waters. The fin identification survey given to nine regional traders demonstrated that they were highly accurate in recognizing pictures of fins from P. glauca and I. oxyrinchus. The overall strong concordance between market categories and fins from single species and the trader accuracy in survey fin identification suggests that monitoring the Chilean fin trade by market names will provide a reasonably accurate picture of the volume of sharks landed by species.     

Knowledge and Practices of Expert Fishermen of South Bahia,Brazil, Regarding the International Shark Fin Market

Little is known about the impacts of the international shark fin trade in emerging economies. This study describes and discusses the knowledge and practices of artisanal fishers in Bahia state, Brazil, concerning the global fin trade. All participants confirmed that fin trade is conducted locally, and 44 (67.7%) admitted to having sold fins. Shark fins reach high prices. Most fishers claim that the number of sharks captured is increasing, and 24.6% (n = 16) claim that this is the reason behind the drop in shark populations. The participants vehemently oppose finning and declare that vessels from other regions catch shark for finning in the area. Fishers believe that surveillance and heavy fines are an effective way to curb finning. Further studies should investigate the sociocultural, economic, and ecological impacts of the sale of fins by artisanal fishers.     

Global-scale genetic identification of hammerhead sharks: Application to assessment of the international fin trade and law enforcement

The future status of sharks is an issue of widespread conservation concern due to declines in many species in the face of high levels of exploitation to satisfy market demands for products, especially fins. Substantial declines in the large-bodied hammerhead sharks, Sphyrna lewini, S. mokarran and S. zygaena, even in regions where some management occurs, indicate that informed conservation measures are warranted for these circumglobally distributed species. Despite the importance of assessing shark catch and trade on a species-specific basis to detect potential overexploitation of individual species, achieving this goal for hammerheads has proven elusive due to difficulties in identification of their products. Here, we present the development and application of a diagnostic, streamlined, five-primer multiplex polymerase chain reaction assay utilizing species-specific primers based on nuclear ribosomal ITS2 for the three hammerhead species throughout their global distribution. Application of this assay to investigations of the fin market confirmed the presence of hammerhead fins in the international trade. A study of the world’s largest fin market in Hong Kong revealed a high concordance between specific Chinese-name trade categories and fins from these three species (“Bai Chun” with S. lewini, “Gui Chun” with S. zygaena and “Gu Pian” with S.␣mokarran), and clear species preferences. This concordance information allows the use of market records for monitoring species-specific trends in trade and exploitation rates. The assay is also proving useful for identification of shark body parts in U.S. fisheries law-enforcement activities. Screening of morphologically identified “ S. lewini” from globally distributed areas using this assay with subsequent whole ITS2 sequencing suggests a cryptic species closely related to S. lewini occurs off the SE USA coast.     

DNA barcoding of traded shark fins,meat and mobulid gill plates in Singapore uncovers numerous threatened species

The shark and ray (Elasmobranchii) trade is a commercially valuable industry that has negative consequences for wild populations. An estimated 100 million sharks are caught each year to supply the demand for cultural cuisines, traditions and practices, including shark fin soup and Traditional Chinese Medicine. Despite the establishment of frameworks and regulations by international trade and conservation bodies as well as national legislations, elasmobranch populations continue to decline. While their conservation becomes an ever more pressing concern, a major obstacle that hampers regulation is the mislabelling and/or misidentification of dried products or carcasses that have had fins removed. Here we use DNA barcoding to identify the species of origin for a variety of shark and ray products readily available to consumers in Singapore, a major importer of these goods. We amplified a fragment of the cytochrome c oxidase subunit I gene from shark fin, cartilage and meat, as well as ray gill plates and meat for DNA sequencing. Our analysis of 207 DNA barcodes yielded 28 positively identified elasmobranch species, eight of which are listed under Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora, and 12 are listed as Endangered or Vulnerable by the International Union for the Conservation of Nature. This information will be useful to regulatory bodies in controlling trade and establishing new or revisiting previous conservation status listings.     

A streamlined, bi-organelle, multiplex PCR approach to species identification: Application to global conservation and trade monitoring of the great white shark, Carcharodon carcharias

The great white shark, Carcharodoncarcharias, is the most widely protectedelasmobranch in the world, and is classified asVulnerable by the IUCN and listed on AppendixIII of CITES. Monitoring of trade in whiteshark products and enforcement of harvest andtrade prohibitions is problematic, however, inlarge part due to difficulties in identifyingmarketed shark parts (e.g., dried fins, meatand processed carcasses) to species level. Toaddress these conservation and managementproblems, we have developed a rapid, moleculardiagnostic assay based on species-specific PCRprimer design for accurate identification ofwhite shark body parts, including dried fins. The assay is novel in several respects: Itemploys a multiplex PCR assay utilizing bothnuclear (ribosomal internal transcribed spacer2) and mitochondrial (cytochrome b) locisimultaneously to achieve a highly robustmeasure of diagnostic accuracy; it is verysensitive, detecting the presence of whiteshark DNA in a mixture of genomic DNAs from upto ten different commercially fished sharkspecies pooled together in a single PCR tube;and it successfully identifies white shark DNAfrom globally distributed animals. Inaddition to its utility for white shark trademonitoring and conservation applications, thishighly streamlined, bi-organelle, multiplex PCRassay may prove useful as a general model forthe design of genetic assays aimed at detectingbody parts from other protected and threatenedspecies.     

Global estimates of shark catches using trade records from commercial markets

Despite growing concerns about overexploitation of sharks, lack of accurate, species-specific harvest data often hampers quantitative stock assessment. In such cases, trade studies can provide insights into exploitation unavailable from traditional monitoring. We applied Bayesian statistical methods to trade data in combination with genetic identification to estimate by species, the annual number of globally traded shark fins, the most commercially valuable product from a group of species often unrecorded in harvest statistics. Our results provide the first fishery-independent estimate of the scale of shark catches worldwide and indicate that shark biomass in the fin trade is three to four times higher than shark catch figures reported in the only global data base. Comparison of our estimates to approximated stock assessment reference points for one of the most commonly traded species, blue shark, suggests that current trade volumes in numbers of sharks are close to or possibly exceeding the maximum sustainable yield levels.     

DNA Barcoding of Shark Meats Identify Species Composition and CITES-Listed Species from the Markets in Taiwan

Background

An increasing awareness of the vulnerability of sharks to exploitation by shark finning has contributed to a growing concern about an unsustainable shark fishery. Taiwan’s fleet has the 4th largest shark catch in the world, accounting for almost 6% of the global figures. Revealing the diversity of sharks consumed by Taiwanese is important in designing conservation plans. However, fins make up less than 5% of the total body weight of a shark, and their bodies are sold as filets in the market, making it difficult or impossible to identify species using morphological traits.

Methods

In the present study, we adopted a DNA barcoding technique using a 391-bp fragment of the mitochondrial cytochrome oxidase I (COI) gene to examine the diversity of shark filets and fins collected from markets and restaurants island-wide in Taiwan.

Results

Amongst the 548 tissue samples collected and sequenced, 20 major clusters were apparent by phylogenetic analyses, each of them containing individuals belonging to the same species (most with more than 95% bootstrap values), corresponding to 20 species of sharks. Additionally, Alopias pelagicus, Carcharhinus falciformis, Isurus oxyrinchus, and Prionace glauca consisted of 80% of the samples we collected, indicating that these species might be heavily consumed in Taiwan. Approximately 5% of the tissue samples used in this study were identified as species listed in CITES Appendix II, including two species of Sphyrna, C. longimanus and Carcharodon carcharias.

Conclusion

DNA barcoding provides an alternative method for understanding shark species composition when species-specific data is unavailable. Considering the global population decline, stock assessments of Appendix II species and highly consumed species are needed to accomplish the ultimate goal of shark conservation.     

Shark fishing in fin soup

Shark DNA extracted from products used in trade(e.g. soup and dried fins) was amplified usingPCR. A strategy is described that permits theidentification of amplified material to species(for mtDNA Cytb and NADH2 loci) and thespecies-specific amplification of basking sharkDNA from mixed templates (for the Cytb locus). These methods will be useful for forensicapplications to govern trade in these heavilyexploited species.     

Composition of Glycosaminoglycans in Elasmobranchs including Several Deep-Sea Sharks: Identification of Chondroitin/Dermatan Sulfate from the Dried Fins of Isurus oxyrinchus and Prionace glauca

Shark fin, used as a food, is a rich source of glycosaminoglyans (GAGs), acidic polysaccharides having important biological activities, suggesting their nutraceutical and pharmaceutical application. A comprehensive survey of GAGs derived from the fin was performed on 11 elasmobranchs, including several deep sea sharks. Chondroitin sulfate (CS) and hyaluronic acid (HA) were found in Isurus oxyrinchus, Prionace glauca, Scyliorhinus torazame, Deania calcea, Chlamydoselachus anguineus, Mitsukurina owatoni, Mustelus griseus and Dasyatis akajei, respectively. CS was only found from Chimaera phantasma, Dalatias licha, and Odontaspis ferox, respectively. Characteristic disaccharide units of most of the CS were comprised of C- and D-type units. Interestingly, substantial amount of CS/dermatan sulfate (DS) was found in the dried fin (without skin and cartilage) of Isurus oxyrinchus and Prionace glauca. ¹H-NMR analysis showed that the composition of glucuronic acid (GlcA) and iduronic acid (IdoA) in shark CS/DS was 41.2% and 58.8% (Isurus oxyrinchus), 36.1% and 63.9% (Prionace glauca), respectively. Furthermore, a substantial proportion of this CS/DS consisted of E-, B- and D-type units. Shark CS/DS stimulated neurite outgrowth of hippocampal neurons at a similar level as DS derived from invertebrate species. Midkine and pleiotrophin interact strongly with CS/DS from Isurus oxyrinchus and Prionace glauca, affording K_d values of 1.07 nM, 6.25 nM and 1.70 nM, 1.88 nM, respectively. These results strongly suggest that the IdoA-rich domain of CS/DS is required for neurite outgrowth activity. A detailed examination of oligosaccharide residues, produced by chondroitinase ACII digestion, suggested that the IdoA and B-type units as well as A- and C-type units were found in clusters in shark CS/DS. In addition, it was discovered that the contents of B-type units in these IdoA-rich domain increased in a length dependent manner, while C- and D-type units were located particularly in the immediate vicinity of the IdoA-rich domain.     

Ecological Drivers of Shark Distributions along a Tropical Coastline

As coastal species experience increasing anthropogenic pressures there is a growing need to characterise the ecological drivers of their abundance and habitat use, and understand how they may respond to changes in their environment. Accordingly, fishery-independent surveys were undertaken to investigate shark abundance along approximately 400 km of the tropical east coast of Australia. Generalised linear models were used to identify ecological drivers of the abundance of immature blacktip Carcharhinus tilstoni/Carcharhinus limbatus, pigeye Carcharhinus amboinensis, and scalloped hammerhead Sphyrna lewini sharks. Results indicated general and species-specific patterns in abundance that were characterised by a range of abiotic and biotic variables. Relationships with turbidity and salinity were similar across multiple species, highlighting the importance of these variables in the functioning of communal shark nurseries. In particular, turbid environments were especially important for all species at typical oceanic salinities. Mangrove proximity, depth, and water temperature were also important; however, their influence varied between species. Ecological drivers may promote spatial diversity in habitat use along environmentally heterogeneous coastlines and may therefore have important implications for population resilience.     

Big catch,little sharks: Insight into Peruvian small‐scale longline fisheries

Shark take, driven by vast demand for meat and fins, is increasing. We set out to gain insights into the impact of small‐scale longline fisheries in Peru. Onboard observers were used to document catch from 145 longline fishing trips (1668 fishing days) originating from Ilo, southern Peru. Fishing effort is divided into two seasons: targeting dolphinfish (Coryphaena hippurus; December to February) and sharks (March to November). A total of 16,610 sharks were observed caught, with 11,166 identified to species level. Of these, 70.6% were blue sharks (Prionace glauca), 28.4% short‐fin mako sharks (Isurus oxyrinchus), and 1% were other species (including thresher (Alopias vulpinus), hammerhead (Sphyrna zygaena), porbeagle (Lamnus nasus), and other Carcharhinidae species (Carcharhinus brachyurus, Carcharhinus falciformis, Galeorhinus galeus). Mean ± SD catch per unit effort of 33.6 ± 10.9 sharks per 1000 hooks was calculated for the shark season and 1.9 ± 3.1 sharks per 1000 hooks were caught in the dolphinfish season. An average of 83.7% of sharks caught (74.7% blue sharks; 93.3% mako sharks) were deemed sexually immature and under the legal minimum landing size, which for species exhibiting k‐selected life history traits can result in susceptibility to over exploitation. As these growing fisheries operate along the entire Peruvian coast and may catch millions of sharks per annum, we conclude that their continued expansion, along with ineffective legislative approaches resulting in removal of immature individuals, has the potential to threaten the sustainability of the fishery, its target species, and ecosystem. There is a need for additional monitoring and research to inform novel management strategies for sharks while maintaining fisher livelihoods.     

Microstructural morphology in early dermal denticles of hammerhead sharks (Elasmobranchii: Sphyrnidae) and related taxa

Mello, W.C., de Carvalho, J.J., Brito, P.M.M. 2011. Microstructural morphology in early dermal denticles of hammerhead sharks (Elasmobranchii: Sphyrnidae) and related taxa. —Acta Zoologica (Stockholm) 00 : 1–7. This study uses scanning electron microscopies to investigate and describe the microstructural diversity of dermal denticles in the family Sphyrnidae, which comprises all living hammerhead shark species, comparing them to other related taxa (i.e. Carcharhinus dussumieri, Carcharhinus plumbeus, Carcharhinus acronotus, Rhizoprionodon acutus, Negaprion brevirostris and Hemigaleus microstoma). The results reveal that sphyrnids present noticeable microstructures in the dermal denticles, distinguishing them from the other related species investigated. Additionally, scale patterns are the same in three distinct body regions (i.e. cephalic, branchial and dorsal fin). Species of Sphyrnidae that reach bigger total lengths and that are widely distributed (i.e. Sphyrna lewini and Sphyrna mokarran) presented more, smaller and nearly hexagonal microstructures that do not cover the entire scale surface, unlike species reaching smaller sizes and restricted to coastal habits (i.e. Sphyrna tiburo, Sphyrna tudes, Sphyrna media and Eusphyra blochii). The sphyrnid scales are similar to R. acutus and C. dussumieri rather than to the other species, but it is not possible to identify the sphyrnid species only by scale features. It is clear that a similar morphology of scales is not necessarily related to similar life habits, and that they are candidates to provide new characters in phylogenetical studies among sphyrnids.     

The Real maccoyii: Identifying Tuna Sushi with DNA Barcodes – Contrasting Characteristic Attributes and Genetic Distances

Background

The use of DNA barcodes for the identification of described species is one of the least controversial and most promising applications of barcoding. There is no consensus, however, as to what constitutes an appropriate identification standard and most barcoding efforts simply attempt to pair a query sequence with reference sequences and deem identification successful if it falls within the bounds of some pre-established cutoffs using genetic distance. Since the Renaissance, however, most biological classification schemes have relied on the use of diagnostic characters to identify and place species.

Methodology/Principal Findings

Here we developed a cytochrome c oxidase subunit I character-based key for the identification of all tuna species of the genus Thunnus, and compared its performance with distance-based measures for identification of 68 samples of tuna sushi purchased from 31 restaurants in Manhattan (New York City) and Denver, Colorado. Both the character-based key and GenBank BLAST successfully identified 100% of the tuna samples, while the Barcode of Life Database (BOLD) as well as genetic distance thresholds, and neighbor-joining phylogenetic tree building performed poorly in terms of species identification. A piece of tuna sushi has the potential to be an endangered species, a fraud, or a health hazard. All three of these cases were uncovered in this study. Nineteen restaurant establishments were unable to clarify or misrepresented what species they sold. Five out of nine samples sold as a variant of “white tuna” were not albacore (T. alalunga), but escolar (Lepidocybium flavorunneum), a gempylid species banned for sale in Italy and Japan due to health concerns. Nineteen samples were northern bluefin tuna (T. thynnus) or the critically endangered southern bluefin tuna (T. maccoyii), though nine restaurants that sold these species did not state these species on their menus.

Conclusions/Significance

The Convention on International Trade Endangered Species (CITES) requires that listed species must be identifiable in trade. This research fulfills this requirement for tuna, and supports the nomination of northern bluefin tuna for CITES listing in 2010.

–The question is not what you look at, but what you see Thoreau

     

Long-Term Changes in Species Composition and Relative Abundances of Sharks at a Provisioning Site

Diving with sharks, often in combination with food baiting/provisioning, has become an important product of today’s recreational dive industry. Whereas the effects baiting/provisioning has on the behaviour and abundance of individual shark species are starting to become known, there is an almost complete lack of equivalent data from multi-species shark diving sites. In this study, changes in species composition and relative abundances were determined at the Shark Reef Marine Reserve, a multi-species shark feeding site in Fiji. Using direct observation sampling methods, eight species of sharks (bull shark Carcharhinus leucas, grey reef shark Carcharhinus amblyrhynchos, whitetip reef shark Triaenodon obesus, blacktip reef shark Carcharhinus melanopterus, tawny nurse shark Nebrius ferrugineus, silvertip shark Carcharhinus albimarginatus, sicklefin lemon shark Negaprion acutidens, and tiger shark Galeocerdo cuvier) displayed inter-annual site fidelity between 2003 and 2012. Encounter rates and/or relative abundances of some species changed over time, overall resulting in more individuals (mostly C. leucas) of fewer species being encountered on average on shark feeding dives at the end of the study period. Differences in shark community composition between the years 2004–2006 and 2007–2012 were evident, mostly because N. ferrugineus, C. albimarginatus and N. acutidens were much more abundant in 2004–2006 and very rare in the period of 2007–2012. Two explanations are offered for the observed changes in relative abundances over time, namely inter-specific interactions and operator-specific feeding protocols. Both, possibly in combination, are suggested to be important determinants of species composition and encounter rates, and relative abundances at this shark provisioning site in Fiji. This study, which includes the most species from a spatially confined shark provisioning site to date, suggests that long-term provisioning may result in competitive exclusion among shark species.     

Internal morphology and function of paired fins in the epaulette shark,hemiscyllium ocellatum

Paired fins and associated internal structures of the epauletic sharkHemiscyllium ocellatum, were described on the basis of three specimens. A comparison with other genera showed the epaulette shark to be, characterized by two elongated basal cartilages articulating with a distally projecting articular condyle on the coracoid, a loosely separated radial series with an intermediate series, a levator pectoralis inferior muscle and an anterolaterally developed depressor pectoralis muscle in the pectoral fin, and an elongated anterior pelvic basal cartilage articulating with a distally projecting articular condyle and an anterolaterally developed depressor pelvicus muscle in the pelvic fin. In captivity, the sharks exhibited both upright and crawling behavior on the bottom by using the pectoral and pelvic fins and bending the body. The distinctive morphological characters are shared by otherHemiscyllium species and are suggested as important factors enabling their unique behavior associated with a complex coral reef habitat.     

Genetic profiling reveals illegal international trade in fins of the great white shark, <Emphasis Type="Italic">Carcharodon carcharias</Emphasis>

Great white sharks are protected by national legislation in several countries, making this species the most widely protected elasmobranch in the world. Although the market demand for shark fins in general has continued to grow, the value and extent of utilization of white shark fins in trade has been controversial. We combine law enforcement with genetic profiling to demonstrate that illegal trade in fins of this species is occurring in the contemporary international market. Furthermore, we document the presence of fins from very young white sharks in the trade, suggesting a multiple-use market (food to trophies) exists for fins of this species. The presence of small fins in the trade contradicts the view that white shark fins have market value only as large display trophies, and not as food. Our findings indicate that effective conservation of protected shark species will require international management regimes that include monitoring of the shark fishery and trade on a species-specific basis.     

Promise and Challenge of DNA Barcoding in Venus Slipper (Paphiopedilum)

Orchidaceae are one of the largest families of flowering plants, with over 27,000 species described and all orchids are listed in CITES. Moreover, the seedlings of orchid species from the same genus are similar. The objective of DNA barcoding is rapid, accurate, and automated species identification, which may be used to identify illegally traded endangered species from vegetative specimens of Paphiopedilum (Venus slipper), a flagship group for plant conservation with high ornamental and commercial values. Here, we selected eight chloroplast barcodes and nrITS to evaluate their suitability in Venus slippers. The results indicate that all tested barcodes had no barcoding gap and the core plant barcodes showed low resolution for the identification of Venus slippers (18.86%). Of the single-locus barcodes, nrITS is the most efficient for the species identification of the genus (52.27%), whereas matK + atpF-atpH is the most efficient multi-locus combination (28.97%). Therefore, we recommend the combination of matK + atpF-atpH + ITS as a barcode for Venus slippers. Furthermore, there is an upper limit of resolution of the candidate barcodes, and only half of the taxa with multiple samples were identified successfully. The low efficiency of these candidate barcodes in Venus slippers may be caused by relatively recent speciation, the upper limit of the barcodes, and/or the sampling density. Although the discriminatory power is relatively low, DNA barcoding may be a promising tool to identify species involved in illegal trade, which has broad applications and is valuable for orchid conservation.