Cytochrome C Oxidase Subunit I Barcode Research Articles

Exploring the Potential in Combining the Two Molecular Approaches, COI Barcoding and PCR-RFLP (COIBar-RFLP) in Identification of Selected Species of the Family Carangidae

The family Carangidae is considered a markedly diverse, widespread taxon. Due to the characteristic “cryptic diversity” and “hybrid speciation” within the family, there is an exigency for taxonomic approaches that go beyond traditional phenotypic modus in identifying species. Mitochondrial cytochrome c oxidase subunit - I (COI) barcoding gene region plays a significant role over phenotypic characters in identifying species. This study evaluates the combination of the two molecular approaches: the COI DNA barcoding and PCR-RFLP for judicious species discrimination. The partial mitochondrial COI gene region of the selected Carangid fish species was amplified, sequenced and confirmed sequences with a mean length of 655 bp were submitted to the main databases; NCBI and BOLD. Intraspecific and interspecific nucleotide divergences were computed by the Kimura-2-Parameter (K2P) and they ranged between (0.00–2.96 %) and (6.46–21.83 %), respectively. The possibility of acquiring the same RFLP profiles given by the restriction enzymes HaeIII and MbOII was observed by the theoretical cleavage of 250 reference sequences of the corresponding gene. Hence, major and minority composite haplotypes of each species were obtained based on the fragment types derived by both HaeIII and MbOII. The resulting divergence values were compatible with the previously reported values for marine fish species. All the species were clearly differentiated by both RFLP banding patterns and the highest probability assumption of getting the same RFLP profile was compatible with the most abundant composite haplotype of each species. This reveals the practicability of the combination of two consolidated molecular approaches, COI barcoding and PCR-RFLP (COIBar-RFLP).

Wing geometric morphometrics and COI barcoding of Culex pipiens subgroup in the Republic of Korea

Two members of the Culex pipiens subgroup, Culex pallens and Culex pipiens f. molestus, are known to occur in the Republic of Korea (ROK). These species exhibit morphologically similar features and are challenging to distinguish below the species level. Therefore, this study utilized wing geometric morphometrics (GM) on the right wing of the Culex pipiens subgroup, alongside sequencing of the cytochrome c oxidase subunit I (COI) region. Mosquitoes were collected from 11 locations between June and October (2020–2022) to minimize regional and seasonal variations. Additionally, Culex pipiens f. pipiens, which is not native to the ROK, was included in the analysis. Culex tritaeniorhynchus, Aedes albopictus, and Anopheles sinensis, the primary vectors in the ROK, were used as outgroups for comparison. All three taxa in the Culex pipiens subgroup could be identified with an 82.4%–97.0% accuracy using GM. However, a comparison of the COI regions of the Culex pipiens subgroup revealed no clear differences between the taxa. These data can be used for accurate identification, contributing to effective mosquito control, in addition to providing a foundation for evolutionary and ecological studies on wing shape differences.

DNA barcoding of black flies (Diptera: Simuliidae) in Indonesia

BackgroundDNA barcoding is a valuable taxonomic tool for rapid and accurate species identification and cryptic species discovery in black flies. Indonesia has 143 nominal species of black flies, but information on their biological aspects, including vectorial capacity and biting habits, remains underreported, in part because of identification problems. The current study represents the first comprehensive DNA barcoding of Indonesian black flies using mitochondrial cytochrome c oxidase subunit I (COI) gene sequences.MethodsGenomic DNA of Indonesian black fly samples were extracted and sequenced, producing 86 COI sequences in total. Two hundred four COI sequences, including 118 GenBank sequences, were analysed. Maximum likelihood (ML) and Bayesian inference (BI) trees were constructed and species delimitation analyses, including ASAP, GMYC and single PTP, were performed to determine whether the species of Indonesian black flies could be delineated. Intra- and interspecific genetic distances were also calculated and the efficacy of COI sequences for species identification was tested.ResultsThe DNA barcodes successfully distinguished most morphologically distinct species (> 80% of sampled taxa). Nonetheless, high maximum intraspecific distances (3.32–13.94%) in 11 species suggested cryptic diversity. Notably, populations of the common taxa Simulium (Gomphostilbia) cheongi, S. (Gomphostilbia) sheilae, S. (Nevermannia) feuerborni and S. (Simulium) tani in the islands of Indonesia were genetically distinct from those on the Southeast Asian mainland (Malaysia and Thailand). Integrated morphological, cytogenetic and nuclear DNA studies are warranted to clarify the taxonomic status of these more complex taxa.ConclusionsThe findings showed that COI barcoding is a promising taxonomic tool for Indonesian black flies. The DNA barcodes will aid in correct identification and genetic study of Indonesian black flies, which will be helpful in the control and management of potential vector species.Graphical abstract

Mitochondrial COI based molecular identification of harvester termite, Anacanthotermes ochraceus (Burmeister, 1839) in Riyadh Region, the Kingdom of Saudi Arabia

ObjectiveTermites are well known for being the most destructive pests of household commodities as well as agricultural crops around the globe. The termite fauna (Isoptera) has about 2650 described species worldwide. Several species are the pests of crops and cause damage to wood structures. MethodsIn the present study, 29 specimens of termites collected from different localities of the Riyadh region were identified using mitochondrial gene cytochrome c oxidase subunit I (COI) deoxyribonucleic acid (DNA) sequence. COI gene was PCR amplified using universal primers (LCO 1490 and HCO 2198). MEGA7 software was used for phylogenetic tree construction which showed that all 29 specimens grouped together in a single clade indicated close relatedness of all specimens. ResultsAll the obtained sequences were submitted into Genbank database and accession numbers were obtained. Phylogenetic analysis showed that all specimens of present research grouped together into a single monophyletic clade, were confirmed to be highly closely related to one another, and proved to be members of the same species. Pairwise nucleotide sequence divergence analysis showed that there was less divergence among all specimens ranging from 0% to 7.8%. Sequence analysis revealed the confirmed precise identification of 29 samples of Anacanthotermes ochraceus with COI barcode analysis. ConclusionsMolecular data analysis has confirmed morphological identification of all 29 studied samples of A. ochraceus. However, this technology offers strong support for identification of cryptic species which are difficult to identify on the basis of morphological features. Further studies of complete mitogenome can be helpful for accurate identification of termites at species level.

Comparison of two molecular barcodes for the study of equine strongylid communities with amplicon sequencing.

Basic knowledge on the biology and epidemiology of equine strongylid species still needs to be improved to contribute to the design of better parasite control strategies. Nemabiome metabarcoding is a convenient tool to quantify and identify species in bulk samples that could overcome the hurdle that cyathostomin morphological identification represents. To date, this approach has relied on the internal transcribed spacer 2 (ITS-2) of the ribosomal RNA gene, with a limited investigation of its predictive performance for cyathostomin communities. Using DNA pools of single cyathostomin worms, this study aimed to provide the first elements to compare performances of the ITS-2 and a cytochrome c oxidase subunit I (COI) barcode newly developed in this study. Barcode predictive abilities were compared across various mock community compositions of two, five and 11 individuals from distinct species. The amplification bias of each barcode was estimated. Results were also compared between various types of biological samples, i.e., eggs, infective larvae or adults. Bioinformatic parameters were chosen to yield the closest representation of the cyathostomin community for each barcode, underscoring the need for communities of known composition for metabarcoding purposes. Overall, the proposed COI barcode was suboptimal relative to the ITS-2 rDNA region, because of PCR amplification biases, reduced sensitivity and higher divergence from the expected community composition. Metabarcoding yielded consistent community composition across the three sample types. However, imperfect correlations were found between relative abundances from infective larvae and other life-stages for Cylicostephanus species using the ITS-2 barcode. While the results remain limited by the considered biological material, they suggest that additional improvements are needed for both the ITS-2 and COI barcodes.

Diversity and genetic connectivity of heteropod (Pterotracheoidea) gastropods in the Tropical Eastern Pacific

AbstractPrevious research focusing on pelagic gastropods in the open ocean has demonstrated that many morphospecies comprise multiple distinct clades or cryptic species that can be distinguished with DNA sequence data. To date, the genetic diversity of the pelagic gastropod fauna of the tropical East Pacific, especially in shallow coastal waters, remains largely unexplored. To document the diversity of pterotracheoids (formerly heteropods) from the coastal waters of the Bay of Panama, we collected, photographed and sequenced fragments of mitochondrial cytochrome c oxidase subunit I (COI) and 16S ribosomal DNA for 60 atlantids, 3 carinariids and 6 pterotracheids. In addition to the COI barcode, our results include the first published 16S sequences for these groups. We found 11 operational taxonomic units (OTUs): 9 in the genus Atlanta, 1 Carinaria and 1 Firoloida. We report the presence of Atlanta oligogyra (Clades A and B), Atlanta turriculata, Atlanta lesueurii, Atlanta helicinoidea (Clade B), Atlanta plana, Atlanta echinogyra, Atlanta inflata and Atlanta frontieri through comparisons of our sequences with previously published sequences. We did not find Atlanta gaudichaudi, Atlanta inclinata, Atlanta tokiokai, Atlanta gibbosa, Atlanta peronii, or Oxygyrus inflatus, which have previously been reported from the region. Haplotype networks and estimates of ΦST illustrate how some species show population differentiation across the tropical Indo‐Pacific region, whereas others show little apparent population structure. For example, the most common haplotypes of A. inflata and of A. turriculata occur in the Indian Ocean, the Central and West Pacific and the tropical East Pacific, whereas individuals of A. frontieri from the Indian Ocean do not share haplotypes with individuals of A. frontieri from the Pacific Ocean. Analyses were limited by sample sizes, but these data suggest that population genetics approaches may be useful for reconstructing population histories of these important, but overlooked, components of the plankton.

DNA Barcoding of Fish Species Diversity in Guizhou, China

Guizhou is an important ecological barrier in the upper reaches of the Yangtze River and the Pearl River basins with abundant fish species. However, fish from these regions are threatened by anthropogenic activities, including overfishing and habitat destruction. Here, we assessed the fish diversity including more than half of the species from the region using DNA barcoding (partial sequence of cytochrome c oxidase subunit I (COI) gene). We obtained 800 mitochondrial COI barcode sequences from 82 genera, 18 families and 8 orders of fishes. The average Kimura two-parameter (K2P) distances within species and genera were 0.35% and 5.44%, respectively. The average interspecific distance was 15.54 times higher than the mean intraspecific distance. Moreover, DNA barcodes revealed 175 operational taxonomic units (OTUs) based on consensus demarcation schemes. Barcoding gaps were detected in 94.81% of morphospecies. Three fish species (Schistura fasciolata, Vanmanenia pingchowensis, and Misgurnus dabryanus) have considerable intraspecific variability, and each was divided into multiple molecular operational taxonomic units (MOTUs) using molecular definition methods (Automatic Barcode Gap Discovery, Refined Single Linkage, General Mixed Yule Coalescent, and Poisson Tree Processes), possibly indicating the occurrence of cryptic species. Altogether, our study reveals the complex diversity of fish species in Guizhou Province, serving as a reference for the conservation and monitoring of fish species in this region.

Integrating morphology and DNA barcodes for identification of Delia sanctijacobi Bigot 1885 (Diptera: Anthomyiidae): new host and new records in South America

Delia sanctijacobi is critically assessed and given a revised description using data from scanning electronic microscopy (SEM) and DNA barcode analysis. This species is recorded for the first time in Brazil and Peru. We provide a morphological identification key (with figures) for Delia species from Brazil, a molecular identification based on COI (cytochrome C oxidase subunit I) barcode sequences and an updated distributional map. We also report the first occurrence of D. sanctijacobi feeding on Brassica species in Brazil. This potential pest was observed in broccoli roots (Brassica oleracea var. italica; Brassicaceae) in União da Vitória, Paraná, southern Brazil, in August and September of 2017. The infested plants displayed reduced growth due to damage to the stem base or death if severely attacked.

Molecular identification of museum preserved type specimens of fish species using DNA barcoding

Type specimens of organisms provide the basis for the identification of species. Proper phenotypic data supported by genetic evidence is crucial for every type of specimen keeping in view its significance. Several methods of species identification both morphological and genetic are being used. DNA barcoding using a fragment of cytochrome c oxidase subunit I (COI) mitochondrial gene is gaining popularity because of its accuracy and efficiency. In this study, six type specimens of endemic fishes from Pakistan preserved at Stephenson Natural History Museum, GC University, Lahore, Pakistan were analyzed for their genetic diversity from other members of the genus. COI barcode sequences of Clupisoma naziri, Barilius vagra pakistanicus, Nemacheilus naziri, Nemacheilus griffithi hazarensis, Schizothorax skarduensis, and Naziritor zhobensis were obtained and analyzed. The obtained sequences were approximately 655bp long. The average Kimura-twoparameter( K2P) distances from other members of genera were 0.608%, 0.44%, 0.42%, 0.608%, 0.945%, and 1.364% for Clupisoma naziri, Barilius vagra pakistanicus, Nemacheilus naziri, Nemacheilus griffithi hazarensis, Schizothorax skarduensis, and Naziritor zhobensis respectively. The nodes in K2P distance-basedNJ (neighbor-joining) trees were supported by high bootstrap values (100%) in all the species. We conclude that COI sequencing provides an effective way of species identification and barcode generation for fish specimens.

Molecular Characterization of Anopheles algeriensis Theobald, 1903 (Diptera: Culicidae) Populations from Europe.

Anopheles algeriensis Theobald, 1903, considered a competent vector of Plasmodium parasites, is a mosquito species widely distributed in the Mediterranean area but rare in Northern and Central Europe. The disappearance of its suitable breeding sites in Italy is having a detrimental effect on the occurrence of this species once common along the Southern coasts and on the islands. Recently, molecular investigations have renewed interest in this species, highlighting a genetic heterogeneity among European populations. In this study, An. algeriensis populations from Italy, Germany, Romania, and Sweden were analyzed by molecular typing of the intergenic transcribed spacer 2 (ITS2). The mitochondrial cytochrome c oxidase subunit I (COI) was also analyzed from specimens collected in Southern Italy. With the aim of investigating the population structure of this species, the obtained data were compared to all publicly available ITS2 and COI sequences of An. algeriensis, adding specimens from Spain and Portugal. The analyses of both markers indicate a split between Iberian populations (Spain for ITS2 and Spain/Portugal for COI) and those from the rest of Europe, revealing two cryptic species. The analysis of the COI barcode revealed a third clade representing a cryptic species present in Danube Delta (Romania). The high levels of genetic divergence among the clades of An. algeriensis indicate that this taxon represents a species complex, potentially harboring several distinct cryptic species.

Egg masses and larval development of the Antarctic cephalaspidean snail Waegelea antarctica (Cephalaspidea: Antarctophilinidae), with notes on egg masses of the related Antarctophiline alata

ABSTRACT We describe, for the first time, egg masses and larval developmental mode of a recently described Antarctic philinoid snail, Waegelea antarctica. Egg masses resembled the gelatinous, attached masses of many temperate philinoid species and contained very large offspring that hatched as developmentally advanced veligers with many juvenile features. Like other Antarctic heterobranch egg masses, development in the masses of W. antarctica appeared to be largely synchronous despite low internal oxygen levels. Hatched larvae could both swim and crawl, and we did not observe metamorphosis over several days. Molecular barcoding using cytochrome c oxidase subunit I (COI) showed an almost perfect (<0.002% difference) match between our specimens from McMurdo Sound in the Ross Sea and a single sequence from a specimen collected >8,000 km away in the Weddell Sea, suggesting either high realized larval dispersal or a recent range expansion. We also describe the egg mass of the related Antarctophiline alata (identified using COI barcoding) from the Ross Sea, which differed from published descriptions in having considerably smaller embryos.

Hundreds of new DNA barcodes for South African sponges

DNA barcoding based on a fragment of the cytochrome c oxidase subunit I (COI) gene from the mitochondrial genome is widely applied in species identification, species discovery and biodiversity studies. The aim of this study was to establish a barcoding reference database of sponges collected from South Africa, and evaluate the applicability of the COI gene for aiding in the identification of sponges in combination with tentative morphological identifications. A total of 317 mitochondrial COI barcode sequences, with an additional 21 extended COI fragments and 24 nuclear ITS sequences, were obtained from 11 orders, 38 families, 58 genera and 124 species of spiculated sponges. A Neighbour Joining (NJ) trees that were reconstructed using these sequences in most cases clustered species in accordance with their current taxonomic identification, and we conclude that COI sequencing can be used to aid in the identification of sponge species. We further demonstrate that DNA barcoding analysis has potential to uncover cryptic sponge species, and to reveal dubious morphological identifications. We recommend that future taxonomic studies of South African sponges incorporate multiple sources of information for species identification or discovery.

Molecular characterization of marine and coastal fishes of Bangladesh through DNA barcodes.

This study describes the molecular characterization of marine and coastal fishes of Bangladesh based on the mitochondrial cytochrome c oxidase subunit I (COI) gene as a marker. A total of 376 mitochondrial COI barcode sequences were obtained from 185 species belonging to 146 genera, 74 families, 21 orders, and two classes of fishes. The mean length of the sequences was 652 base pairs. In Elasmobranchii (Sharks and rays), the average Kimura two parameter (K2P) distances within species, genera, families, and orders were 1.20%, 6.07%, 11.08%, and 14.68%, respectively, and for Actinopterygii, the average K2P distances within species, genera, families, and orders were 0.40%, 6.36%, 14.10%, and 24.07%, respectively. The mean interspecies distance was 16‐fold higher than the mean intraspecies distance. The K2P neighbor‐joining (NJ) trees based on the sequences generally clustered species in accordance with their taxonomic position. A total of 21 species were newly recorded in Bangladesh. High efficiency and fidelity in species identification and discrimination were demonstrated in the present study by DNA barcoding, and we conclude that COI sequencing can be used as an authentic identification marker for Bangladesh marine fish species.

Revealing the biodiversity of Chilean birds through the COI barcode approach.

The mitochondrial cytochrome c oxidase subunit I (COI) gene is an effective molecular tool for the estimation of genetic variation and the identification of bird species. This molecular marker is used to differentiate among Chilean bird species by analyzing barcodes for 76 species (197 individuals), comprising 28 species with no previous barcode data and 48 species with sequences retrieved from the BOLD and GenBank databases. The DNA barcodes correctly identified 94.7% of the species analyzed (72 of 76 species). Mean intraspecific K2P distance was 0.3% (range 0–8.7%). Within the intraspecific divergence range, three species, Phrygilus gayi, Sephanoides sephanoides and Curaeus curaeus, showed relatively high intraspecific divergence (1.5–8.7%), possibly due to the presence of a species complex or geographic isolation of sub-populations. Mean interspecific K2P distance was 24.7% (range 1.3–43.5%). Consequently, the intraspecific K2P distance showed limited overlap with interspecific K2P distance. The mean intraspecific divergence in our study was similar to that found in temperate regions of South America (0.24%). However, it was approximately one order of magnitude lower than values reported for bird species in tropical regions of northern South America (1.8–2.13%). This result suggests that bird species from Chile show low levels of genetic structure and divergence. The small overlap between intra- and inter-specific distances implies that COI barcodes could be used as an effective tool to identify nearly all the Chilean bird species analyzed.

The efficacy of COI barcoding and ISSR markers in molecular identification of diverse bird Sternula saundersi populations along the Red Sea coast, Kingdom of Saudi Arabia

Aim: To demonstrate the first DNA barcode and molecular diversity of bird Sternula saundersi (Saunders’s tern) populations collected from the Red Sea coast at the Kingdom of Saudi Arabia. Methodology: The mitochondrial cytochrome c oxidase subunit I (COI) gene was isolated and sequenced from 26 different individuals of S. saundersi, whose blood samples were collected from two different areas at the Red Sea coast of the Kingdom of Saudi Arabia. Also, the genetic diversity of 26 individuals were investigated using highly polymorphic nuclear markers, i.e., inter simple sequence repeat [ISSR] technique. Results: The COI gene sequences of all 26 individuals were documented and submitted to the GenBank database. Five unique single nucleotide polymorphisms were found in all individuals of S. saundersi that were not observed in other species of the genus Sternula. Consequently, these five nucleotides can be used as a distinctive DNA fingerprint of S. saundersi. Additionally, it was possible to identify molecular diversity within different populations from two different areas using eight different ISSR markers. Interpretation: The present data demonstrate the power of COI marker gene in identifying S. saundersi, i.e., it can be utilized to discriminate between different species of the genus Sternula. In addition, highly polymorphic nuclear markers (ISSR) were demonstrated to have significant power to accurately distinguish between individuals collected from different widely spaced areas.

Exposing the Barcoding Void: An Integrative Approach to Study Snail-Borne Parasites in a One Health Context.

Trematodes are snail-borne parasites of major zoonotic importance that infect millions of people and animals worldwide and frequently hybridize with closely related species. Therefore, it is desirable to study trematodiases in a One Health framework, where human and animal trematodes are considered equally important. It is within this framework that we set out to study the snail and trematode communities in four artificial lakes and an abattoir in Zimbabwe. Trematode infections in snails were detected through multiplex PCR protocols. Subsequently, we identified snails by sequencing a partial mitochondrial cytochrome c oxidase subunit I (COI) fragment, and trematodes (adults from the abattoir and larval stages detected in snails) using COI and nuclear rDNA markers. Of the 1,674 collected snails, 699 were molecularly analyzed, in which we identified 12 snail and 19 trematode species. Additionally, three parasite species were sampled from the abattoir. Merely four trematode species were identified to species level through COI-based barcoding. Moreover, identification of members of the superfamilies Opisthorchioidea and Plagiorchioidea required a phylogenetic inference using the highly conserved 18S rDNA marker, as no related COI reference sequences were present in public databases. These barcoding challenges demonstrate a severe barcoding void in the available databases, which can be attributed to the neglected status of trematodiases. Adding to this, many available sequences cannot be used as different studies use different markers. To fill this gap, more studies on African trematodes, using a standardized COI barcoding region, are desperately needed.

Efficient COI barcoding using high throughput single-end 400\u2009bp sequencing

BackgroundOver the last decade, the rapid development of high-throughput sequencing platforms has accelerated species description and assisted morphological classification through DNA barcoding. However, the current high-throughput DNA barcoding methods cannot obtain full-length barcode sequences due to read length limitations (e.g. a maximum read length of 300 bp for the Illumina’s MiSeq system), or are hindered by a relatively high cost or low sequencing output (e.g. a maximum number of eight million reads per cell for the PacBio’s SEQUEL II system).ResultsPooled cytochrome c oxidase subunit I (COI) barcodes from individual specimens were sequenced on the MGISEQ-2000 platform using the single-end 400 bp (SE400) module. We present a bioinformatic pipeline, HIFI-SE, that takes reads generated from the 5′ and 3′ ends of the COI barcode region and assembles them into full-length barcodes. HIFI-SE is written in Python and includes four function modules of filter, assign, assembly and taxonomy. We applied the HIFI-SE to a set of 845 samples (30 marine invertebrates, 815 insects) and delivered a total of 747 fully assembled COI barcodes as well as 70 Wolbachia and fungi symbionts. Compared to their corresponding Sanger sequences (72 sequences available), nearly all samples (71/72) were correctly and accurately assembled, including 46 samples that had a similarity score of 100% and 25 of ca. 99%.ConclusionsThe HIFI-SE pipeline represents an efficient way to produce standard full-length barcodes, while the reasonable cost and high sensitivity of our method can contribute considerably more DNA barcodes under the same budget. Our method thereby advances DNA-based species identification from diverse ecosystems and increases the number of relevant applications.

Description of Sarcocystis scandentiborneensis sp. nov. from treeshrews (Tupaia minor, T. tana) in northern Borneo with annotations on the utility of COI and 18S rDNA sequences for species delineation.

Sarcocystis scandentiborneensis sp. nov. was discovered in histological sections of striated musculature of treeshrews (Tupaia minor, T. tana) from Northern Borneo. Sarcocysts were cigar-shaped, 102 μm–545 μm long, and on average 53 μm in diameter. The striated cyst wall varied in thickness (2–10 μm), depending on whether the finger-like, villous protrusions (VP) were bent. Ultrastructurally, sarcocysts were similar to wall type 12 but basal microtubules extended into VPs that tapered off with a unique U-shaped, electron-dense apical structure. In phylogenetic trees of the nuclear 18S rRNA gene, S. scandentiborneensis formed a distinct branch within a monophyletic subclade of Sarcocystis spp. with (colubrid) snake-rodent life cycle. We mapped all intraspecific (two haplotypes) and interspecific nucleotide substitutions to the secondary structure of the 18S rRNA gene: in both cases, the highest variability occurred within helices V2 and V4 but intraspecific variability mostly related to transitions, while transition/transversion ratios between S. scandentiborneensis, S. zuoi, and S. clethrionomyelaphis were skewed towards transversions. Lack of relevant sequences restricted phylogenetic analysis of the mitochondrial Cytochrome C oxidase subunit I (COI) gene to include only one species of Sarcocystis recovered from a snake host (S. pantherophisi) with which the new species formed a sister relationship. We confirm the presence of the functionally important elements of the COI barcode amino acid sequence of S. scandentiborneensis, whereby the frequency of functionally important amino acids (Alanine, Serine) was markedly different to other taxa of the Sarcocystidae. We regard S. scandentiborneensis a new species, highlighting that structurally or functionally important aspects of the 18S rRNA and COI could expand their utility for delineation of species. We also address the question why treeshrews, believed to be close to primates, carry a parasite that is genetically close to a Sarcocystis lineage preferably developing in the Rodentia as intermediate hosts.

Molecular diversity of freshwater fishes of Bangladesh assessed by DNA barcoding

This study represents the comprehensive molecular identification of freshwater fishes of Bangladesh based on a fragment of the cytochrome c oxidase subunit I (COI) gene in the mitochondrial genome. A total of 315 mitochondrial COI barcode sequences were obtained from 153 species,114 genera, 49 families and 16 orders of fishes. The mean length of the sequences was 652 base pairs. For all the samples, %G was significantly lower compared to the other nucleotides and %GC was lower compared to %AT (p-value ˂ 0.05). Also, a significantly lower %GC content was observed in second and third codon position compared to the first one in all the samples (1st>2nd>3rd, p-value˂ 0.05). The average K2P distances within species, genera, families and orders were 0.38%, 7.02%, 12.75% and 18.68%, respectively. The mean interspecific distance was 18-fold higher than the mean intraspecific distance. The K2P neighbor-joining (NJ) trees based on the sequences generally clustered species according to their taxonomic position. A total of 12 species were newly recorded in Bangladesh. High efficiency in species identification were demonstrated in the present study by DNA barcoding, and concluded that COI sequencing can be used as an authentic identification marker for freshwater fish species.
 Bangladesh J. Zool. 48(1): 1-19, 2020

DNA barcoding for identification of marine gastropod species from Hainan island, China

Morphological identification of gastropods can be difficult considering the larva of species and high similarity among the same genera. DNA barcoding has been widely used in species identification and biodiversity research. The purpose of this study was to explore the feasibility of using the cytochrome c oxidase subunit I (COI) gene for the identification of gastropod species and to construct a reliable DNA barcoding reference database of gastropods in Hainan island, China. A total of 306 mitochondrial COI barcode sequences were obtained from 120 species, 35 families and 7 orders of gastropods. The average length of the sequence was 640 base pairs. The average genetic distances based on Kimura two parameter (K2P) within species, genera, families, orders and classes were 0.9 %, 14.7 %, 18.9 %, 24.5 % and 28.6 %, respectively. Most of the gastropod species could be identified using COI sequences. Our results confirmed that the identification method combining morphology and DNA barcode greatly improved the efficiency of species identification. In this study, we found three new record species in China, namely Semiricinula tissoti (Petit de la Saussaye, 1852), Engina alveolata (Kiener, 1836) and Wallaconchis ater (Lesson, 1831). Overall, this study revealed that the identification of gastropods by DNA barcoding is efficient, and COI sequencing technology can be used for the identification of gastropod species and thereby can be used to manage fisheries and assess biodiversity.