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The correct identification of mosquito species is important for effective mosquito vector control. However, the standard morphological identification of mosquito species based on the available keys is not easy with specimens in the field due to missing or damaged morphological features during mosquito collections, often leading to the misidentification of morphologically indistinguishable. To resolve this problem, we collected mosquito species across Thailand to gather genetic information, and evaluated the DNA barcoding efficacy for mosquito species identification in Thailand. A total of 310 mosquito samples, representing 73 mosquito species, were amplified using mitochondrial cytochrome c oxidase subunit I (COI) primers. The average maximum intraspecific genetic variation of the 73 mosquito species was 1% ranged from 0–5.7%. While, average minimum interspecific genetic variation (the distance to the nearest neighbour) of the 73 mosquito species was 7% ranged from 0.3–12.9%. The identification of success rates based on the "Best Match," "Best Close Match," and "All Species Barcodes" methods were 97.7%, 91.6%, and 81%, respectively. Phylogenetic analyses of Anopheles COI sequences demonstrated a clear separation between almost all species (except for those between An. baimaii and An. dirus), with high bootstrap support values (97%–99%). Furthermore, phylogenetic analyses revealed potential sibling species of An. annularis, An. tessellatus, and An. subpictus in Thailand. Our results indicated that DNA barcoding is an effective molecular approach for the accurate identification of mosquitoes in Thailand. [ABSTRACT FROM AUTHOR]

Muscid flies are of great medical and veterinary importance because of their ability to transmit diseases to humans and animals. Their larvae are found on dead bodies and are used as a forensic indicator for estimating Post-Mortem Interval (PMI). A drawback of morphological identification of these larvae is lack of an identification key which covers all the life stages of the taxa. To overcome the problem, entomologists now focus on the alternative methods, which can speed up the identification process. Barcoding region of Cytochrome oxidase I (COI)l is believed to be the best among other markers for precise identification and to mark out phylogeny. During the present study, total 63 specimens of 24 species belonging to 7 genera representing the various muscid subfamilies were used to check the efficacy of COI gene in identification. 11 species were collected from various regions (Punjab, Haryana, Himachal Pradesh, Uttarakhand) of North India during the years 2015–2019, and sequences of rest of the species were downloaded from the database of Genbank. The percentage of intraspecific divergences ranged from 0%-0.5% for species of India and 0%-3.0% for the sequences from other countries. The interspecific divergence ranged from 3.1% to 17.3% and 3.3% to 16.7% respectively for Indian species and species from the other part of the world. 1000 bootstrapped Neighbor-joining with Kimura-2 parameter phylogenetic tree constructed based on COI gene sequences, clearly separated 24 species of these Muscid flies. This is the first ever study on COI gene based identification of medico-legal, veterinary and forensically important Muscid flies from India. [ABSTRACT FROM AUTHOR]

Two unresolved questions in marine population connectivity are: (1) the relative importance of contemporary and historical factors (e.g., ongoing gene flow versus past demographic events), and (2) to what extent species subject to the same evolutionary forces exhibit similar phylogeographic patterns. Here, we address these questions using two species from New Zealand's rocky shore that have very similar distributions and life histories, but very different larval dispersal abilities: the cat's eye snail Lunella smaragda has short-lived pelagic larvae (3–4 days) while the half-crab Petrolisthes elongatus has a longer pelagic larval duration (3–4 weeks). A large number of individuals of these species were collected (n = 727 and 440) at different locations (31 and 20) throughout their wide New Zealand distribution. These species were analysed for both mitochondrial DNA cytochrome oxidase I (COI) and nuclear ribosomal internal transcribed spacer (ITS-1) variation. Contrary to expectations, the species with much greater dispersal potential, P. elongatus, exhibited much greater population differentiation (> fivefold for mtDNA ΦST, > 50-fold for nDNA ΦST). This study highlights that species along the same coast can show remarkably different patterns of population structure, and that although there appear to be some common geographic discontinuities in New Zealand, there are few common overall patterns that apply to many species. The study reinforces the observation that predictions of population structure based on life history are often not upheld, and shows that differences in demographic history may be an important factor in driving contemporary patterns of genetic diversity. [ABSTRACT FROM AUTHOR]

The habitats of Galago moholi are suspected to be largely fragmented, while the species is thought to be expanding further into the southernmost fringe of its range, as well as into human settlements. To date, no intraspecific molecular genetic studies have been published on G. moholi. Here we estimate the genetic diversity and connectivity of populations of G. moholi using two mitochondrial gene regions, the cytochrome C oxidase subunit I gene (COI) and the displacement loop of the control region (D-loop). Samples from five localities in northern South Africa were obtained from archived collections. The two mitochondrial DNA gene regions were amplified and sequenced to provide population summary statistics, differentiation [proportion of the total genetic variation in a population relative to the total genetic variance of all the populations (FST), differentiation within populations among regions (ΦST)], genetic distance and structure. There was discernible genetic structure among the individuals, with two COI and six D-loop haplotypes belonging to two genetically different groups. There was population differentiation among regions (FST = 0.670; ΦST = 0.783; P < 0.01). However, there were low levels of differentiation among populations, as haplotypes were shared between distant populations. Adjacent populations were as divergent from each other as from distant populations. The results suggest that genetic introgression, most likely due to past migrations or recent unintentional translocations that include the animal trade, may have led to connectivity among populations. [ABSTRACT FROM AUTHOR]

Purpose: Taenia multiceps resides in the small intestine of carnivores such as dogs, foxes, woles, jackals, while Coenurus cerebralis which is the larval form usually settle in the central nervous system and spinal cord of intermediate hosts like sheep, cattle and goats. The aim of this study was to analyze the haplotype diversity, genetic variation and population structure of the mt-CO1 gene sequences of sheep isolates of T. multiceps had been submitted to GenBank from different countries. Methods: A total of 102 sequences from the mt-CO1 gene fragment belong to T. multiceps sheep isolates were used for bioinformatic analyses. Haplotype analysis, phylogenetic analysis and diversity, neutrality, fixation and gene flow analyses were applied to the sequences. Results: As a result, 20 haplotypes together with different multiple nucleotide changes were determined after the sequence analysis. Trimmed fragment length was 337 bp hereby 19 polymorphic areas, 12 of which were parsimony informative, were identified, and any insertion–deletion was found. The number of mutations between major haplotypes and the others range from one to nine. The highest (0.72) genetic differentiation (Fst value) was observed between Turkey and Egypt populations while the lowest (− 0.22) was reported from Greece. These findings are important in terms of showing the diversity of nucleotide variation in T. multiceps sheep isolates. Conclusions: This study serves as the basis for future large-scale studies on T. multiceps worldwide epidemiology, bioecology, geographic distribution and population structure. [ABSTRACT FROM AUTHOR]

Determining the genetic structure and population dynamics of harvested species are key aspects of effective fisheries management. New Zealand scampi (Metanephrops challengeri) is a species of nethropid lobster that supports a commercial fishery on the continental shelf and slope around New Zealand. To determine the genetic structure and historical population demography of New Zealand scampi, we sequenced a 623 base-pair fragment of the mitochondrial gene cytochrome c oxidase subunit one (CO1) from 321 individuals across six widely separated sites off the coast of mainland New Zealand and the subantarctic Auckland Islands, representing much of the species' distribution. The Auckland Islands population was found to be genetically distinct from all other sampled populations, potentially resulting from an absence of continuous suitable habitat between the central-eastern South Island and the Auckland Islands. An isolation-by-distance pattern of genetic structure was also detected. With the exception of the Mercury Islands, all sampled populations showed signs of recent population expansion, potentially linked to changes in habitat availability during the glacial-interglacial cycles of the Pleistocene. We recommend that the current NZ scampi management areas and harvesting rates for each remain unchanged. [ABSTRACT FROM AUTHOR]

The scallop Argopecten purpuratus is a heavily exploited resource along the coasts of Peru and north-central Chile, especially after El Niño events, when the species undergoes high increases in abundance. Little is known about its genetic structure or demographic history, two important factors to ensure sustainable exploitation. We sequenced the cytochrome oxidase I and cytochrome b genes of 116 individuals from six localities (between 05°44′S 80°53′W and 23°31′S 70°33′W). We found high levels of genetic diversity in the analyzed populations. No geographical structuring was observed in the haplotype network, which consisted of a few central, widely distributed haplotypes, and many derived population-specific haplotypes separated by few mutations. This pattern suggests a recent population expansion and moderate to low current gene flow among populations. Mismatch analysis, neutrality tests, and a Bayesian skyline analysis confirmed the occurrence of a past event of population expansion approximately 5,000 years ago, which coincides with increasingly stronger and more frequent El Niño events. [ABSTRACT FROM AUTHOR]

In the present study, DNA barcoding was used to assess the percentage accuracy of morphological base identification of spiders from the agriculture fields of district Layyah, Punjab, Pakistan. A total of 872 spiders were captured from June to August of 2017. All the collected spiders were brought to molecular laboratory at GC University Lahore, preserved in 95% ethanol and stored at −20 °C until the DNA extraction. Spiders were evaluated morphologically on the basis of different identification Keys and Catalogs. Morphological identification revealed the presence of 12 families, 29 genra and 49 species. To evaluate the authenticity of morphological identification, tissue samples of 96 specimens were sent to Canadian Center for Biodiversity and Genomics, University of Guelph, Canada. A 658-base pair sequence of COI (Cytochrome c Oxidase Subunit I) of 90 specimens was retrieved successfully, which confirmed the presence of 11 families, 25 genra and 47 species. On the basis of molecular results, all the misidentified specimens were then allotted the correct taxon. Overall accuracy of morphological based identification was 88%. It is concluded from the present study that morphological investigations to identify a spider, are satisfactory but to enhance the accuracy, pace and credibility of results, molecular technique like DNA barcoding is considerable. Furthermore, to magnify authenticity of evaluation of spiders, integrated barcoding- combination of molecular methods and conventional taxonomy- is compulsory. [ABSTRACT FROM AUTHOR]

We evaluated the extent of intraspecific and interspecific genetic distances for two highly diverse infraorders of Odonata: Anisoptera and Zygoptera. All cytochrome c oxidase subunit I sequences (cox1), the region chosen for zoological DNA barcoding, present in GenBank for each infraorder were downloaded and curated. For Anisoptera, the final dataset consisted of 2,961 individual cox1 sequences for 536 species and the equivalent numbers for Zygoptera were 2,477 sequences for 497 species. More than 7 million individual genetic comparisons were made and the results indicated that there is a tendency towards a barcoding gap, but that the size of the gap may not be sufficient to robustly infer identities for some taxa. DNA barcoding may be of less use for some odonate taxa, perhaps pertaining to misidentifications in global databases. However, at local scales or with more confined taxonomical sampling, this tool may yet be beneficial in identifying these charismatic organisms. [ABSTRACT FROM AUTHOR]

Identification of fish species have so far been carried out mostly by classical morpho-taxonomy. In the present study, however, an attempt has been taken to identify two species of fishes Ulua mentalis and Pinjalo pinjalo of order Perciformes which happens to be the first record in Odisha coast Bay of Bengal, India during the year 2015, using DNA barcoding technique for reconfirmation over conventional morpho-taxonomy. During recent past, study of molecular-taxonomical profile of mitochondrial DNA in general and Cytochrome Oxidase subunit I (COI) gene in particular has gained enormous importance for accurate identification of species. In the present study, the partial COI sequence of Ulua mentalis and Pinjalo pinjalo were generated. Analysis using the COI gene produced phylogenetic trees in concurrence with other multi gene studies and we came across the identical phylogenetic relationship considering Neighbor-Joining and Maximum Likelihood tree. Moreover, these molecular data set further testified in Bayesian framework to reevaluate the exact taxonomic groupings within the family. Surprisingly, Ulua mentalis and Pinjalo pinjalo seems to be closely related to their sister taxa. [ABSTRACT FROM AUTHOR]

Amphipods constitute an abundant part of Icelandic deep-sea zoobenthos yet knowledge of the diversity of this fauna, particularly at the molecular level, is scarce. The present work aims to use molecular methods to investigate genetic variation of the Amphipoda sampled during two IceAGE collecting expeditions. The mitochondrial cytochrome oxidase subunit 1 (COI) of 167 individuals originally assigned to 75 morphospecies was analysed. These targeted morhospecies were readily identifiable by experts using light microscopy and representative of families where there is current ongoing taxonomic research. The study resulted in 81 Barcode Identity Numbers (BINs) (of which >90% were published for the first time), while Automatic Barcode Gap Discovery revealed the existence of 78 to 83 Molecular Operational Taxonomic Units (MOTUs). Six nominal species (Rhachotropis helleri, Arrhis phyllonyx, Deflexilodes tenuirostratus, Paroediceros propinquus, Metopa boeckii, Astyra abyssi) appeared to have a molecular variation higher than the 0.03 threshold of both p-distance and K2P usually used for amphipod species delineation. Conversely, two Oedicerotidae regarded as separate morphospecies clustered together with divergences in the order of intraspecific variation. The incongruence between the BINs associated with presently identified species and the publicly available data of the same taxa was observed in case of Paramphithoe hystrix and Amphilochus manudens. The findings from this research project highlight the necessity of supporting molecular studies with thorough morphology species analyses. [ABSTRACT FROM AUTHOR]

Molecular genetics has proven to be an essential tool for studying the ecology, evolution and epidemiology of parasitic nematodes. However, research effort across nematode taxa has not been equal and biased towards nematodes parasitic in vertebrates. We characterize the evolutionary genetics of the mermithid nematode Thaumamermis zealandica Poinar, 2002 and its host, the sandhopper Bellorchestia quoyana (Milne-Edwards, 1840) (Talitridae: Amphipoda), across sandy beaches of New Zealand's South Island. We test the hypothesis that parasite population genetic structure mirrors that of its host. Sandhoppers and their parasites were sampled at 13 locations along the island's southeastern coast. Sequencing of the mitochondrial gene cytochrome c oxidase subunit 1 (CO1) from B. quoyana reveals a regional pattern of population structure that suggests a northward pattern of dispersal. Surprisingly, no population structure was observed for T. zealandica. In fact, sequencing of three commonly used markers revealed no intraspecific parasite variation. This result suggests that mermithid mtDNA may evolve at an extraordinarily slow pace, perhaps as a result of extensive and frequent changes in gene order and mitochondrial genome length. Furthermore, a mermithid phylogeny based on sequences of the 18S and 28S ribosomal RNA genes suggests that a systematic revision of the family is necessary. [ABSTRACT FROM AUTHOR]

Biodiversity is well defined with the assistance of taxonomy, the science which classifies organisms with “species” as the basic unit. Presently, in taxonomy, morphological type specimen is complimented with its molecular data from “type” gene sequences and species status is granted. For this, DNA barcoding using standardized DNA gene(s) is performed in which Cytochrome c oxidase I gene of mitochondrial DNA is referred as the primary barcode region for animal kingdom. Being a popular mitochondrial marker, this gene is reported to possess two barcode regions with limited overlap, “Folmer” and “Palumbi” regions particularly in decapod crustaceans. Here we demonstrate the issues originated as a result of incorporation of both these barcode regions together in addressing various aspects of DNA barcoding like specimen identification, population analysis, and phylogeny in decapod crustaceans using COI sequences (“Folmer” and “Palumbi”) ofMacrobrachium rosenbergiias reference sequences. [ABSTRACT FROM PUBLISHER]

Daphnia is a freshwater zooplankton species with controversial taxonomy due to its high morphological variation linked to environmental factors and inter-specific hybridization and polyploidy in some groups. The aim of the present study is to examine molecular diversity of some Daphnia species in Turkey and to establish DNA barcodes of Turkish Daphnia species. Sequence analysis was performed using 540 bp region of cytochrome oxidase subunit I gene of mitochondrial DNA. A total of 34 haplotypes have been identified for Turkey. Daphnia pulex complex was divided into two clades with 16.1% sequence divergence according to molecular taxonomy based on Kimura 2-parameter. The clade which was molecularly diverged from Daphnia pulex with 16.1% sequence divergence was found to show 99% similarity with Daphnia cf. pulicaria (sensu Alonso 1996) instead of Daphnia pulicaria Forbes, 1893. Furthermore, this study has contributed to Turkish zoogeography by demonstrating the distribution of Daphnia species in Turkey. [ABSTRACT FROM AUTHOR]

Host-parasite co-evolution can lead to genetic differentiation among isolated host-parasite populations and local adaptation between parasites and their hosts. However, tests of local adaptation rarely consider multiple fitness-related traits although focus on a single component of fitness can be misleading. Here, we concomitantly examined genetic structure and co-divergence patterns of the trematode Coitocaecum parvum and its crustacean host Paracalliope fluviatilis among isolated populations using the mitochondrial cytochrome oxidase I gene ( COI). We then performed experimental cross-infections between two genetically divergent host-parasite populations. Both hosts and parasites displayed genetic differentiation among populations, although genetic structure was less pronounced in the parasite. Data also supported a co-divergence scenario between C. parvum and P. fluviatilis potentially related to local co-adaptation. Results from cross-infections indicated that some parasite lineages seemed to be locally adapted to their sympatric (home) hosts in which they achieved higher infection and survival rates than in allopatric (away) amphipods. However, local, intrinsic host and parasite characteristics (host behavioural or immunological resistance to infections, parasite infectivity or growth rate) also influenced patterns of host-parasite interactions. For example, overall host vulnerability to C. parvum varied between populations, regardless of parasite origin (local vs. foreign), potentially swamping apparent local co-adaptation effects. Furthermore, local adaptation effects seemed trait specific; different components of parasite fitness (infection and survival rates, growth) responded differently to cross-infections. Overall, data show that genetic differentiation is not inevitably coupled with local adaptation, and that the latter must be interpreted with caution in a multi-trait context. [ABSTRACT FROM AUTHOR]

DNA barcoding using a fragment of the mitochondrial cytochrome c oxidase subunit 1 gene (COI) has proven to be successful for species-level identification in many animal groups. However, most studies have been focused on relatively small datasets or on large datasets of taxonomically high-ranked groups. We explore the quality of DNA barcodes to delimit species in the diverse chironomid genus Tanytarsus (Diptera: Chironomidae) by using different analytical tools. The genus Tanytarsus is the most species-rich taxon of tribe Tanytarsini (Diptera: Chironomidae) with more than 400 species worldwide, some of which can be notoriously difficult to identify to species-level using morphology. Our dataset, based on sequences generated from own material and publicly available data in BOLD, consist of 2790 DNA barcodes with a fragment length of at least 500 base pairs. A neighbor joining tree of this dataset comprises 131 well separated clusters representing 121 morphological species of Tanytarsus: 77 named, 16 unnamed and 28 unidentified theoretical species. For our geographically widespread dataset, DNA barcodes unambiguously discriminate 94.6% of the Tanytarsus species recognized through prior morphological study. Deep intraspecific divergences exist in some species complexes, and need further taxonomic studies using appropriate nuclear markers as well as morphological and ecological data to be resolved. The DNA barcodes cluster into 120–242 molecular operational taxonomic units (OTUs) depending on whether Objective Clustering, Automatic Barcode Gap Discovery (ABGD), Generalized Mixed Yule Coalescent model (GMYC), Poisson Tree Process (PTP), subjective evaluation of the neighbor joining tree or Barcode Index Numbers (BINs) are used. We suggest that a 4–5% threshold is appropriate to delineate species of Tanytarsus non-biting midges. [ABSTRACT FROM AUTHOR]

Evaluating species boundaries remains a significant challenge in rare, difficult to collect, and/or understudied groups because of a lack of available data. Uninformative morphology, unknown ecologies and/or geographical distributions, inadequate comparative sequence data, and limited sample sizes present substantial challenges when applying commonly used methods for species delimitation. Here we present an approach that overcomes the challenges previously mentioned by integrating phylogeny, genetic distances, and a fixed diagnostic character (i.e. colour pattern) into the species delimitation process. The genus Entomobrya ( Collembola) includes many species marked by complex and variable colour patterns. Many Entomobrya species have been named based exclusively on colour pattern, but the value of this character as a species-level diagnostic marker has been challenged. To test the hypothesis that colour forms in Entomobrya represent independent evolutionary lineages, i.e. distinct species, we used phylogenetic methods to evaluate the association between colour pattern and molecular variation in the cytochrome c oxidase I gene ( COI) in 11 species of North American Entomobrya. The comparative analysis focused on 13 colour forms distributed amongst the species Entomobrya assuta, Entomobrya clitellaria, Entomobrya ligata, and Entomobrya quadrilineata. Phylogenetic analysis and genetic divergences sorted the 13 colour forms into seven independent evolutionary lineages, including three morphologically cryptic lineages diagnosable by colour pattern. However, genetic divergence did not always correlate with colour pattern variation, indicating that the diagnostic utility of colour pattern is species dependent and requires individual evaluation for each species. We propose that incorporation of the explicit species delimitation criteria developed for this study will result in a substantial advance in the identification and description of species in understudied taxa. © 2015 The Linnean Society of London [ABSTRACT FROM AUTHOR]

The deep ocean supports a highly diverse and mostly endemic fauna, yet little is known about how or where new species form in this remote ecosystem. How speciation occurs is especially intriguing in the deep sea because few obvious barriers exist that would disrupt gene flow. Geographic and bathymetric patterns of genetic variation can provide key insights into how and where new species form. We quantified the population genetic structure of a protobranch bivalve, Neilonella salicensis, along a depth gradient (2200-3800 m) in the western North Atlantic using both nuclear (28 S and calmodulin intron) and mitochondrial (cytochrome c oxidase subunit I) loci. A sharp genetic break occurred for each locus between populations above 2800 m and below 3200 m, defining two distinct clades with no nuclear or mitochondrial haplotypes shared between depth regimes. Bayesian phylogenetic analyses provided strong support for two clades, separated by depth, within N. salicensis. Although no morphological divergence was apparent, we suggest that the depth-related population genetic and phylogenetic divergence is indicative of a cryptic species. The frequent occurrence of various stages of divergence associated with species formation along bathymetric gradients suggests that depth, and the environmental gradients that attend changes in depth, probably play a fundamental role in the diversification of marine organisms, especially in deep water. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113, 897-913. [ABSTRACT FROM AUTHOR]

The role of dispersal potential on phylogeographic structure, evidenced by the degree of genetic structure and the presence of coincident genetic and biogeographic breaks, was evaluated in a macrogeographic comparative approach along the north-central coast of Chile, across the biogeographic transition zone at 30°S. Using 2,217 partial sequences of the mitochondrial Cytochrome Oxidase I gene of eight benthic invertebrate species along ca. 2,600 km of coast, we contrasted dispersal potential with genetic structure and determined the concordance between genetic divergence between biogeographic regions and the biogeographic transition zone at 30°S. Genetic diversity and differentiation highly differed between species with high and low dispersal potential. Dispersal potential, sometimes together with biogeographic region, was the factor that best explained the genetic structure of the eight species. The three low dispersal species, and one species assigned to the high dispersal category, had a phylogeographic discontinuity coincident with the biogeographic transition zone at 30°S. Furthermore, coalescent analyses based on the isolation-with-migration model validate that the split between biogeographic regions north and south of 30°S has a historic origin. The signatures of the historic break in high dispersers is parsimoniously explained by the homogenizing effects of gene flow that have erased the genetic signatures, if ever existed, in high dispersers. Of the four species with structure across the break, only two had significant albeit very low levels of asymmetric migration across the transition zone. Historic processes have led to the current biogeographic and phylogeographic structure of marine species with limited dispersal along the north-central coast of Chile, with a strong lasting impact in their genetic structure. [ABSTRACT FROM AUTHOR]

The amphipod Hyalella azteca is commonly used as a model for determining safe concentrations of contaminants in freshwaters. The authors sequenced the mitochondrial cytochrome c oxidase subunit I ( COI) gene for representatives of 38 populations of this species complex from US and Canadian toxicology research laboratories and eastern North American field sites to determine their genetic relationships. With 1 exception, all US and Canadian laboratory cultures sampled were identified as conspecific. In 22 wild populations spanning 5 US states and 1 Canadian province, the commonly occurring laboratory species was found only in northern Florida, USA. Therefore, the diversity of the H. azteca species complex detected in the wild is not accurately represented in North American laboratories, questioning the reliability of H. azteca cultures currently in use to accurately predict the responses of wild populations in ecotoxicological assays. The authors also examined the utility of different COI nucleotide fragments presently in use to determine phylogenetic relationships in this group and concluded that saturation in DNA sequences leads to inconsistent relationships between clades. Amino acid sequences for COI were not saturated and may allow a more accurate phylogeny estimate. Hyalella azteca is crucial for developing water-quality regulations; therefore, laboratories should know and standardize the strain(s) they use to confidently compare toxicity tests across laboratories and determine whether they are an appropriate surrogate for their regions. Environ Toxicol Chem 2013;32:2637-2647. © 2013 SETAC [ABSTRACT FROM AUTHOR]

Bees ( Apidae), of which there are more than 19 900 species, are extremely important for ecosystem services and economic purposes, so taxon identity is a major concern. The goal of this study was to optimize the DNA barcode technique based on the Cytochrome c oxidase ( COI) mitochondrial gene region. This approach has previously been shown to be useful in resolving taxonomic inconsistencies and for species identification when morphological data are poor. Specifically, we designed and tested new primers and standardized PCR conditions to amplify the barcode region for bees, focusing on the corbiculate Apids. In addition, primers were designed to amplify small COI amplicons and tested with pinned specimens. Short barcode sequences were easily obtained for some Bombus century-old museum specimens and shown to be useful as mini-barcodes. The new primers and PCR conditions established in this study proved to be successful for the amplification of the barcode region for all species tested, regardless of the conditions of tissue preservation. We saw no evidence of Wolbachia or numts amplification by these primers, and so we suggest that these new primers are of broad value for corbiculate bee identification through DNA barcode. [ABSTRACT FROM AUTHOR]

We used a DNA barcoding approach to identify specimens of the Daphnia pulex complex occurring in New Zealand lakes, documenting the establishment of non-indigenous North American Daphnia 'pulex'. Morphological delineation of species in this complex is problematic due to a lack of good morphological traits to distinguish the species, as there is a relatively high degree of morphological stasis within the group through evolutionary time. Accordingly, genetic analyses were used to determine the specific identity and likely geographic origin of this species. Morphologically, individuals most closely resembled Daphnia pulicaria or Daphnia pulex sensu lato, which cannot be separated morphologically. Furthermore, each of these taxa comprises separate species in North America and Europe, despite carrying the same names. We identified individuals using a 658 bp nucleotide portion of the mitochondrial cytochrome c oxidase subunit 1 gene (COI) as North American Daphnia 'pulex', being distinct from European Daphnia pulex sensu stricto and D. pulicaria from Europe or North America. Cellulose allozyme electrophoresis was used to confirm that individuals were not hybrids with D. pulicaria. North American Daphnia 'pulex' in New Zealand were first recorded in New Zealand from South Island lakes that are popular for overseas recreational fishers, indicating a possible source of introduction for this species (e.g. on/in fishing gear). Our study provides an additional example of how genetic techniques can be used for the accurate identification of non-indigenous taxa, particularly when morphological species determination is not possible. The growth of global databases such as GenBank and Barcode of Life Datasystems (BOLD) will further enhance this identification capacity. [ABSTRACT FROM AUTHOR]

The relationship between species diversity and environmental parameters is poorly understood for the mobile macrofauna of deep-sea habitats due to under-sampling and subsequent lack of accurate taxonomic information. To redress this, cytochrome oxidase c subunit I ( COI) DNA sequences were used to estimate species diversity and to compare phoxocephalid amphipod assemblages among 20 stations encompassing a range of environmental conditions. Two regions, east ( Chatham Rise) and west ( Challenger Plateau) of New Zealand were sampled to depths of 200-1200 m with an epibenthic sled. Using a comparison among identified morphospecies, we found a clear gap in sequence divergences between 6% and 13% and used a 6% threshold to designate molecular operational taxonomic units ( MOTUs), as a surrogate to putative species. DNA sequences ( n = 297) revealed high total diversity ( n = 49 MOTUs), as well as high beta diversity (28 MOTUs found at single location only). Novel phoxocephalid MOTUs were found at most stations, especially on Challenger Plateau and the flanks of Chatham Rise. Analyses of interstation assemblages revealed a major split between regions, indicating minimal overlap in taxon distributions. A cluster of highly similar stations was identified, broadly distributed over the crest of Chatham Rise, in association with elevated food availability, probably resulting from higher surface productivity and relatively shallow depth. Accordingly, multivariate analysis revealed a strong correlation between phoxocephalid assemblages and food supply. This study highlights the value of molecular approaches, in particular COI sequences, for quantifying and comparing diversity in under-sampled and/or under-studied taxa. [ABSTRACT FROM AUTHOR]

We evaluated whether cytochrome c oxidase subunit I (COI) barcodes that have been previously suggested for birds are useful for identifying species of the genus Larus, which are resident or migratory birds in Korea. We found 31 intra- or interspecific COI haplotypes from 12 of 13 Larus species in Korea. Haplotype analyses showed that the COI barcodes could not distinguish some interspecific haplotypes from 6 of 12 Larus species because there were no nucleotide substitutions among their COI haplotypes. The neighbor-joining tree formed shallow branches in the clades expected for L. saundersi, L. crassirostris, L. canus, L relictus, and L. ridbundus. In the nine Larus species, COI haplotypes were not grouped as distinct entities that were correctly assigned to their corresponding species, resulting in polytypic clades. These results indicate that the COI sequences need to be cautiously selected as a DNA barcode for identifying species of Korean Larus birds. [ABSTRACT FROM AUTHOR]

A DNA-based barcode identification system that is applicable to all animal species will provide a simple, universal tool for the identification of fish species. The barcode system is based on sequence diversity in subunit 1 cytochrome c oxidase ( COI) gene. Identification and characterization of fish species based on morphological characters are sometimes found to be erroneous and environmentally affected. There are no studies on the genus Ompok in India at molecular level and species identification of the Ompok is usually carried out through morphological features. A total of 106 samples from three species Ompok pabda, O. pabo and O. bimaculatus were collected from eight sampling sites of seven Indian rivers. One hundred and six sequences were generated from COI region of three Ompok species and 21 haplotypes were observed. The sequence analysis of COI gene revealed three genetically distinct Ompok species and exhibited identical phylogenetic resolution among them. The partial COI gene sequence can be used as a diagnostic molecular marker for identification and resolution of taxonomic ambiguity of Ompok species. [ABSTRACT FROM AUTHOR]

Coral reefs are believed to be one of the most diverse ecosystems, but the true magnitude of their biodiversity and patterns of endemism is uncertain. This uncertainty stems partly from the relative paucity of investigations on the small, difficult to collect taxa (cryptofauna) that may make up the majority of reef biodiversity and require specialized expertise for morphological identification. To assess the extent of diversity in some of the reef micro-cryptofauna, we analyzed 414 bp of the mitochondrial cytochrome oxidase subunit I gene from 556 individuals representing two brooding amphipod species ( Leucothoe ashleyae and Leucothoe kensleyi). These amphipods are commensal inside the branching vase sponge Callyspongia vaginalis, and were sampled throughout Florida and the Caribbean. Phylogenetic analyses revealed 11 deeply divergent, strongly supported lineages (seven L. ashleyae and four L. kensleyi) each with very narrow geographic ranges. The level of intraspecific lineage divergence for both morphospecies was among the highest reported for any marine crustacean (12.4-26.0% uncorrected), and exceeded that of congeners from nine diverse amphipod families, as well as the patristic genetic distance suggested as a threshold for crustacean species delineation. These findings suggest a history of cryptic speciation within each morphospecies, concomitant with a pronounced period of morphological stasis involving each of the morphotypes. The observation of multiple, highly divergent, evolutionary significant units, each endemic to Florida and Caribbean island locations, supports the emerging view that coral reef biodiversity, especially in the cryptofaunal component, is likely vastly underestimated. [ABSTRACT FROM AUTHOR]

DNA barcodes have proven invaluable in identifying and distinguishing insect pests, most notably for determining the provenance of exotic invasives, but relatively few insect natural enemies have been barcoded. We used universal invertebrate primers and for Lepidoptera, to amplify 658 bp at the 5′ end of the mitochondrial cytochrome oxidase c subunit I (COI) gene in five species of lady beetles from crop fields in six states in the US Mid-Atlantic, Plains and Midwest: three native species, Hippodamia convergens Guérin-Méneville, H. parenthesis (Say) and Coleomegilla maculata (De Geer); and two exotic species , Harmonia axyridis (Pallas) and Coccinella septempunctata Linnaeus . Sequence divergences within species were low, never exceeding 0.9% (Kimura 2-parameter distances). Sequence divergences between the two Hippodamia species ranged from 14.7 to 16.4%, mirroring the relationships found for other arthropod taxa. Among the exotic species, C. septempunctata sequences were as variable as those of the three native species, while H. axyridis populations comprised a single haplotype. Limited data on two Coleomegilla subspecies, C. m. lengi Timberlake and C. m. fuscilabris (Mulsant), are consistent with their belonging to the same species, although morphological and reproductive data indicate that they represent separate species. Our results support the general utility of COI barcodes for distinguishing and diagnosing coccinellid species, but point to possible limitations in the use of barcodes to resolve species assignments in recently divergent sibling species. [ABSTRACT FROM AUTHOR]

DNA barcoding—sequencing a standard region of the mitochondrial cytochrome c oxidase 1 gene (COI)—promises a rapid, accurate means of identifying animals to a species level. This study establishes that sequence variability in the barcode region permits discrimination of 98% of 201 fish species from the Canadian Pacific. The average sequence variation within species was 0.25%, while the average distance separating species within genera was 3.75%. The latter value was considerably lower than values reported in other studies, reflecting the dominance of the Canadian fauna by members of the young and highly diverse genus Sebastes. Although most sebastids possessed distinctive COI sequences, four species did not. As a partial offset to these cases, the barcode records indicated the presence of a new, broadly distributed species of Paraliparis and the possibility that Paraliparis pectoralis is actually a species pair. The present study shows that most fish species in Pacific Canadian waters correspond to a single, tightly cohesive array of barcode sequences that are distinct from those of any other species, but also highlights some taxonomic issues that need further investigation. [ABSTRACT FROM AUTHOR]

DNA barcoding has gained increased recognition as a molecular tool for species identification in various groups of organisms. In this preliminary study, we tested the efficacy of a 615-bp fragment of the cytochrome c oxidase I (COI) as a DNA barcode in the medically important family Simuliidae, or black flies. A total of 65 (25%) morphologically distinct species and sibling species in species complexes of the 255 recognized Nearctic black fly species were used to create a preliminary barcode profile for the family. Genetic divergence among congeners averaged 14.93% (range 2.83–15.33%), whereas intraspecific genetic divergence between morphologically distinct species averaged 0.72% (range 0–3.84%). DNA barcodes correctly identified nearly 100% of the morphologically distinct species (87% of the total sampled taxa), whereas in species complexes (13% of the sampled taxa) maximum values of divergence were comparatively higher (max. 4.58–6.5%), indicating cryptic diversity. The existence of sibling species in Prosimulium travisi and P. neomacropyga was also demonstrated, thus confirming previous cytological evidence about the existence of such cryptic diversity in these two taxa. We conclude that DNA barcoding is an effective method for species identification and discovery of cryptic diversity in black flies. [ABSTRACT FROM AUTHOR]

Background: DNA barcoding based on the mitochondrial cytochrome oxidase subunit I gene (cox1 or COI) has been successful in species identification across a wide array of taxa but in some cases failed to delimit the species boundaries of closely allied allopatric species or of hybridising sister species. Methodology/Principal Findings: In this study we extend the sample size of prior studies in birds for cox1 (2776 sequences, 756 species) and target especially species that are known to occur parapatrically, and/or are known to hybridise, on a Holarctic scale. In order to obtain a larger set of taxa (altogether 2719 species), we include also DNA sequences of two other mitochondrial genes: cytochrome b (cob) (4614 sequences, 2087 species) and 16S (708 sequences, 498 species). Our results confirm the existence of a wide gap between intra- and interspecies divergences for both cox1 and cob, and indicate that distance-based DNA barcoding provides sufficient information to identify and delineate bird species in 98% of all possible pairwise comparisons. This DNA barcoding gap was not statistically influenced by the number of individuals sequenced per species. However, most of the hybridising parapatric species pairs have average divergences intermediate between intraspecific and interspecific distances for both cox1 and cob. Conclusions/Significance: DNA barcoding, if used as a tool for species discovery, would thus fail to identify hybridising parapatric species pairs. However, most of them can probably still assigned to known species by character-based approaches, although development of complementary nuclear markers will be necessary to account for mitochondrial introgression in hybridising species. [ABSTRACT FROM AUTHOR]

The 655 bp cytochrome c oxidase subunit I barcode region of single specimens of 388 species of fishes (four Holocephali, 61 Elasmobranchii and 323 Actinopterygii) was examined. All but two ( Urolophus cruciatus and Urolophus sufflavus) showed different cox1 nucleotide sequences (99.5% species discrimination); the two that could not be resolved are suspected to hybridize. Most of the power of cox1 nucleotide sequence analysis for species identification comes from the degenerate nature of the genetic code and the highly variable nature of the third codon position of amino acids. Variation at the third codon position is bimodally distributed, and the more variable mode is dominated by amino acids with four or six codons, while the less variable mode is dominated by amino acids with two codons. The ratio of nonsynonymous to synomymous changes is much less than one, indicating that this gene is subject to strong purifying selection. Consequently, cox1 amino acid sequence diversity is much less than nucleotide sequence diversity and has very poor species resolution power. Fourteen of the 16 amino acid residues recognized as having important functions in the region of cox1 sequenced were completely conserved over all 388 species (and the bovine cox1 sequence), with one fish species varying at one of these sites, and three fish at another site. No significant differences in amino acid conservation were observed between residues in helices, strands and turns. Patterns of nucleotide and amino acid variability were very similar between elasmobranchs and actinopterygians. [ABSTRACT FROM AUTHOR]

Morphology-based keys support accurate identification of many taxa. However, identification can be difficult for taxa that are either not well studied, very small, members of cryptic species complexes, or represented by immature stages. For such cases, DNA barcodes may provide diagnostic characters. Ecologists and evolutionary biologists deposit museum vouchers to document the species studied in their research. If DNA barcodes are to be used for identification, then both the DNA and the specimen from which it was extracted should be vouchered. We describe a protocol for the nondestructive extraction of DNA from terrestrial arthropods, using as examples members of the orders Acarina, Araneae, Coleoptera, Diptera, and Hymenoptera chosen to represent the ranges in size, overall sclerotization, and delicacy of key morphological characters in the group. We successfully extracted sequenceable DNA from all species after 1–4 h of immersion in extraction buffer. The extracted carcasses, processed and imaged using protocols standard for the taxon, were distinguishable from closely related species, and adequate as morphological vouchers. We provide links from the carcasses and DNA vouchers to image (MorphBank) and sequence (GenBank) databases. [ABSTRACT FROM AUTHOR]

The article discusses that DNA sequencing can help in identifying species using the barcoding system. The DNA sequencing and barcode has been proposed as a technology that might hasten species identification. The use of mitochondrial cytochrome c oxidase subunit one gene employed a possible DNA marker for species. DNA barcoding resolves most species and provide means of highlighting potential cryptic.

Phragmatopoma spp . are marine, reef-building polychaetes that inhabit the intertidal and shallow subtidal zones of both coasts of the Americas. Phragmatopoma californica is found in the Pacific Ocean along the California coast south to Mexico, while Phragmatopoma caudata inhabits the Caribbean islands and Atlantic Ocean from the Florida coast south to Brazil. Although apparently geographically isolated, P. californica and P. caudata have been found to interbreed (Pawlik ) and thus their specific taxonomic relationship has been unclear. In this study, two genes, cytochrome c oxidase subunit I (COI) and the first internal transcribed spacer region (ITS-1), were sequenced to assess the specific status of P. californica and P. caudata as well as Phragmatopoma virgini. Comparison of sequences revealed that samples of P. californica shared no COI haplotypes or ITS-1 sequences with P. caudata. Phylogenetic analyses, including maximum parsimony and Bayesian methods, clustered each species in separate, well-supported clades with genetic distances between them being greater than between either contested species or the uncontested, valid species, P. virgini. Thus, the molecular data support that P. californica and P. caudata are separate species. However, the sample of individuals of P. virgini included one genetically divergent individual, whose morphology was found to match that of a species formerly recognized as P. moerchi but since synonymized with P. virgini. Divergences among lineages were dated using the COI sequences, after adjustment for non-clock-like behavior. Consequently, we suggest that P. virgini and P. caudata are sister taxons and that P. californica diverged from the P. virgini/P. caudata clade ∼5.7 mya with P. virgini diverging from P. caudata ∼3 mya. [ABSTRACT FROM AUTHOR]

Short DNA sequences from a standardized region of the genome provide a DNA barcode for identifying species. Compiling a public library of DNA barcodes linked to named specimens could provide a new master key for identifying species, one whose power will rise with increased taxon coverage and with faster, cheaper sequencing. Recent work suggests that sequence diversity in a 648-bp region of the mitochondrial gene, cytochrome c oxidase I (COI), might serve as a DNA barcode for the identification of animal species. This study tested the effectiveness of a COI barcode in discriminating bird species, one of the largest and best-studied vertebrate groups. We determined COI barcodes for 260 species of North American birds and found that distinguishing species was generally straightforward. All species had a different COI barcode(s), and the differences between closely related species were, on average, 18 times higher than the differences within species. Our results identified four probable new species of North American birds, suggesting that a global survey will lead to the recognition of many additional bird species. The finding of large COI sequence differences between, as compared to small differences within, species confirms the effectiveness of COI barcodes for the identification of bird species. This result plus those from other groups of animals imply that a standard screening threshold of sequence difference (10× average intraspecific difference) could speed the discovery of new animal species. The growing evidence for the effectiveness of DNA barcodes as a basis for species identification supports an international exercise that has recently begun to assemble a comprehensive library of COI sequences linked to named specimens. Can a single gene be used to identify all different animal species? An analysis of 255 North American birds shows that cytochrome c oxidase I might have this potential. [ABSTRACT FROM AUTHOR]