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Simple Summary: The tribe Dacnusini is a small braconid parasitoid wasp within the subfamily Alysiinae (Hymenoptera: Braconidae), consisting of 31 valid genera and more than 877 species worldwide. Most members are exclusively endoparasitoids of leaf-mining Diptera (Agromyzidae), and several species have been utilized in commercial biological control programs, underscoring their essential role in natural pest management. However, due to low interspecific variability and significant morphological convergence, distinguishing between genera and species within the genera in Dacnusini presents a challenge. In this study, we report three genera, Victorovita Tobias, Coloneura Foerster, and Laotris Nixon, that were discovered in China for the first time. By integrating DNA barcoding and morphological evidence, three new species were identified through multiple species delimitation methods. Additionally, two new species records from China are reported, and identification keys for species of Laotris Nixon, 1943 are provided. Dacnusini is a species-rich tribe in the subfamily Alysiinae, with most species exclusively serving as parasitoids of leaf-mining Diptera (Agromyzidae). The number of genera discovered in China remains limited, which is apparently insufficient considering the global diversity of species and genera within this tribe, particularly given the vast and ecologically diverse landscapes of China. In the present study, three new record genera, Victorovita Tobias, Coloneura Foerster, and Laotris Nixon, were documented for the first time in China. In addition, the species delimitation approach and haplotype network analyses based on the COI sequences, combined with morphological evidence, were employed to delimit species. The findings indicated three new species: Laotris glabella sp. nov., Laotris aethidentata sp. nov., and Victorovita aequalis sp. nov. Additionally, K2P divergences showed no overlap between intra- and interspecific genetic distances in the Laotris and Victorovita species. Detailed descriptions for new species and keys to the species of Laotris and Victorovita are provided in this paper, along with the documentation of two new species records for China: Victorovita caudata (Szépligeti, 1901) and Coloneura stylata Foerster, 1863. [ABSTRACT FROM AUTHOR]

Genetic diversity and ecology are two important aspects of vector species crucial to a full understanding of disease epidemiology. In this study, we examined genetic diversity, genetic structure and the physiochemical parameters of the breeding habitats of the two significant black fly vector species, Simulium chumpornense Takaoka and Kuvangkadilok and S. nodosum Puri, from Laos. Genetic diversity of S. chumpornense in Laos was relatively high with maximum genetic divergence of 3.05% but no significant genetic differentiation between populations. Comparisons with conspecific populations from Thailand also found no genetic differentiation between the two countries. This possibly related to the recent history of this species, as a very recent (31,000 years ago) historical population expansion was detected. Physicochemical parameters of the breeding habitats suggest the ability to utilize diverse stream sizes from small flows (1 m wide) to huge rivers (290 m or more wide). Populations of S. nodosum from Laos had low genetic diversity with maximum genetic divergence of 2.56% and no genetic structuring among populations. Comparisons with those reported from other countries revealed five genetically divergent lineages (I–V) with minimum genetic divergence of 1.36%. The majority (42 of 52) of specimens from Laos belonged to lineage I and the remaining (10) comprised lineage II. Lineage I is the largest and representative of specimens from Thailand, Myanmar, Vietnam, Taiwan and Laos. Population history analysis revealed that lineage I had undergone recent demographic expansion dating back to 7000 years ago. This very recent population expansion resembles others reported from Thailand and possibly indicates a response to increasing human and domestic animals following the agricultural revolution. Breeding sites of S. nodosum are diverse in respect to elevation, velocity, water conductivity and streambed particle size. The ability to utilize a wide variety of breeding sites could promote the production of a large adult population, which can be a pest for humans and other animals. [ABSTRACT FROM AUTHOR]

Astylus atromaculatus Blanchard, 1843 (Coleoptera: Melyridae) is a pollinivorous beetle native to southern South America, which has invaded South Africa more than a century ago. Adults and/or larvae may occasionally damage flowers, seeds, and seedlings of various crops. Severe cattle intoxication has also been reported in Argentina, Uruguay, and South Africa following consumption of alfalfa and forage grasses infested with A. atromaculatus. Despite its economic impact, essential genetic information is lacking for this species. The present paper provides the first DNA barcode reference sequences for A. atromaculatus based on the standard 5’ fragment (658 bp) of the cytochrome c oxidase subunit I gene. The sequences obtained exhibited pairwise distances of ≤ 1.82% among them, and ~ 90% nucleotide identity with the homologous gene fragment in the morphologically similar Astylus variegatus Germar, 1824. The use of this molecular marker to explore the intraspecific variability of A. atromaculatus in central Argentina showed 21 different haplotypes, out of 32 individuals analyzed. A very high haplotype diversity (Hd = 0.962 ± 0.019) and a moderate nucleotide diversity (π = 0.00778 ± 0.00079) were recorded. The haplotype network displayed a diffuse structure due to the abundance of singletons and possible missing haplotypes, with the most common haplotype comprising only 15.6% of the specimens collected. Future research with increased sampling size and geographic coverage will allow for a better understanding of the population genetics of this pest, and consequently, for developing efficient management practices. [ABSTRACT FROM AUTHOR]

ABSTRACT Low coverage ‘genome‐skims’ are often used to assemble organelle genomes and ribosomal gene sequences for cost‐effective phylogenetic and barcoding studies. Natural history collections hold invaluable biological information, yet poor preservation resulting in degraded DNA often hinders polymerase chain reaction‐based analyses. However, it is possible to generate libraries and sequence the short fragments typical of degraded DNA to generate genome‐skims from museum collections. Here we introduce a snakemake toolkit comprised of three pipelines skim2mito, skim2rrna and gene2phylo, designed to unlock the genomic potential of historical museum specimens using genome skimming. Specifically, skim2mito and skim2rrna perform the batch assembly, annotation and phylogenetic analysis of mitochondrial genomes and nuclear ribosomal genes, respectively, from low‐coverage genome skims. The third pipeline gene2phylo takes a set of gene alignments and performs phylogenetic analysis of individual genes, partitioned analysis of concatenated alignments and a phylogenetic analysis based on gene trees. We benchmark our pipelines with simulated data, followed by testing with a novel genome skimming dataset from both recent and historical solariellid gastropod samples. We show that the toolkit can recover mitochondrial and ribosomal genes from poorly preserved museum specimens of the gastropod family Solariellidae, and the phylogenetic analysis is consistent with our current understanding of taxonomic relationships. The generation of bioinformatic pipelines that facilitate processing large quantities of sequence data from the vast repository of specimens held in natural history museum collections will greatly aid species discovery and exploration of biodiversity over time, ultimately aiding conservation efforts in the face of a changing planet. [ABSTRACT FROM AUTHOR]

The soybean pod borer, Leguminivora glycinivorella (Matsumura), is an important tortricid pest species widely distributed in most parts of China and its adjacent regions. Here, we analyzed the genetic diversity and population differentiation of L. glycinivorella using diverse genetic information including the standard cox1 barcode sequences, mitochondrial genomes (mitogenomes), and single‐nucleotide polymorphisms (SNPs) from genotyping‐by‐sequencing. Based on a comprehensive sampling (including adults or larvae of L. glycinivorella newly collected at 22 of the total 30 localities examined) that covers most of the known distribution range of this pest, analyses of 543 cox1 barcode sequences and 60 mitogenomes revealed that the traditionally recognized and widely distributed L. glycinivorella contains two sympatric and widely distributed genetic lineages (A and B) that were estimated to have diverged ∼1.14 million years ago during the middle Pleistocene. Moreover, low but statistically significant correlations were recognized between genetic differentiation and geographic or environmental distances, indicating the existence of local adaptation to some extent. Based on SNPs, phylogenetic inference, principal component analysis, fixation index, and admixture analysis all confirm the two divergent sympatric lineages. Compared with the stable demographic history of Lineage B, the expansion of Lineage A had possibly made the secondary contact of the two lineages probable, and this process may be driven by the climate fluctuation during the late Pleistocene as revealed by ecological niche modeling. [ABSTRACT FROM AUTHOR]

Holoplanktonic organisms are thought to have enormous populations sizes, expansive geographic ranges, and low species diversity. Previous work suggests that pelagic gastropods are no exception to this generalization. However, most of these data are derived from temperate species, whereas the bulk of pteropod diversity occurs in the tropics. Here, we present DNA barcode data for COI focused on the under‐studied limacinoid, gymnosome, and pseudothecosome pteropods collected from the coastal waters of Panama. We applied four molecular species delimitation approaches to determine the number of operational taxonomic units (OTUs) identifiable from our sequences and all data available from Barcode of Life Database (BOLD) and GenBank and compared these to the Barcode Index Number generated by the BOLD. Assemble Species by Automatic Partitioning (ASAP), Poisson Tree Processes (PTP), Bayesian Poisson Tree Processes (bPTP), and Generalized Mixed Yule Coalescent (GMYC) methods generally gave congruent results and suggest that pteropod diversity in the tropics is significantly underrepresented both in DNA sequence databases and in the number of named OTUs. Of 42 specimens collected and sequenced, we recovered 16 OTUs, only one of which belonged to an OTU already represented in sequence databases. Neighbor‐joining trees including the previously published sequences show that in all three groups morphospecies can include genetically divergent lineages. Many very divergent (>15% from nearest neighbor) taxa are also still represented by only a single sequence, suggesting that there is a large amount of cryptic or pseudo‐cryptic diversity still to be described. To aid this future endeavor, we include some preliminary 16S data derived from new pteropod‐specific primers. [ABSTRACT FROM AUTHOR]

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

Background: Thoracophelia Ehlers, 1897 is a genus of Opheliidae characterised by the body divided into three distinct regions, modified parapodia in chaetiger 10 and a ventral groove restricted to the posterior half of the body. To date, 18 species have been described in the genus. Amongst them, six species have been recorded in northeast Asia. New information: A new species, Thoracophelia foliformis sp. nov., was discovered in the intertidal zone of the Yellow Sea, South Korea. This is the first Thoracophelia species report from the Yellow Sea. This new species is closely related to T. dillonensis (Hartman, 1938) from California and T. ezoensis Okuda, 1936 from Japan in having pectinate branchiae. However, the new species can be distinguished from the two species by the unique combination of the following characteristics: 15 pairs of wrinkled pectinate branchiae with 12–15 filaments at best development and a foliaceous mid-ventral plate in the pygidium instead of one or two thick ventral cirri. Detailed descriptions and illustrations of T. foliformis sp. nov. are provided. Sequences of the mitochondrial cytochrome c oxidase subunit I (COI), nuclear 18S ribosomal DNA (rDNA) and 28S rDNA of the new species were determined and analysed. [ABSTRACT FROM AUTHOR]

Background: Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. Methods: Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. Results: The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28–30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74–18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. Conclusions: This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided. [ABSTRACT FROM AUTHOR]

The jumping spider species Gedea pinguis Cao & Li, 2016 is newly recorded from Vietnam and redescribed based on the sole male specimen. A detailed redescription, illustrations of the copulatory organs, and somatic features, along with an updated distribution map are provided. DNA barcode derived from the mitochondrial cytochrome c oxidase subunit I (COI) gene of the species G. pinguis from Vietnam is also included for future use and to compare the genetic distances with Gedea cf. tibialis Zabka, 1985 from Malaysia. With the addition of a new record, the genus Gedea Simon, 1902 in Vietnam now reaches 3 species, but doubts arise regarding the conspecificity of the two opposite sex species, Gedea tibialis Żabka, 1985 (male) and Gedea typica (Żabka, 1985) (female), as they share the same type locality. Additionally, a dichotomous key to all Gedea species is also given in this study. [ABSTRACT FROM AUTHOR]

The Afrotropical Cerurinae (Notodontidae) genera Cerurina Kiriakoff and Afrocerura Kiriakoff are revised. Historically, significant confusion has existed surrounding the identities of members of these 2 closely related genera. Using morphology and genetic data derived from mitochondrial COI barcodes and anchored hybrid enrichment nuclear genomics, we present an integrated taxonomic approach. Upon examining all types, historical material in global collections, and numerous recently collected specimens, we are able to associate available names with the taxa in these genera as well as describe four new species of Afrocerura for which no names were available. Historical misidentifications are also addressed. For nearly all species, both sexes and their genitalia are figured. In total, we recognize one species in Cerurina with 2 subspecies: C. marshalli marshalli Hampson (type species) and C. marshalli cameroona (Bethune-Baker) comb. n. , and 8 species in Afrocerura : A. leonensis (Hampson) (type species), A. takanoi sp. n. , A. bifasciata (Janse), A. inexpectata sp. n. , A. tanganyikae Kiriakoff stat. n. , A. ethiopica sp. n. , A. thomensis (Talbot) and A. smithi sp. n. Color photos of C. m. cameroona larvae are presented for the first time and are compared to larvae of C. m. marshalli. [ABSTRACT FROM AUTHOR]

In our recent examination of the Phintella specimens collected from Hainan Tropical Rainforest National Park, a new species and the unknown female of P. liae Wang, Mi & Peng, 2023 were recognised, based on the morphological characteristics and molecular evidence.A new species of Phintella Strand, 1906 is described: P. hongkan sp. nov. (♂♀) from Hainan, China. The unknown female of P. liae Wang, Mi & Peng, 2023 is also described for the first time. Diagnostic photos of both species are provided.

The genus Carnus Nitzsch, 1818 comprises small ectoparasites that feed on the blood of juvenile avians. They are characterised by dealated adults with setose abdominal intersegmental membranes. Carnus orientalis Maa, 1968 was previously recorded in Malaysia and the Ryukyu Islands of Japan, parasitising two owl species: Ketupa ketupu (Horsfield, 1821) and Otus elegans (Cassin, 1852). This study confirms the occurrence of C. orientalis in Taiwan and presents a new host record, along with COI barcode sequences. Additionally, the study also elucidates the difficulties posed by blood meal contamination and pseudogene amplification as confounding factors intrinsic to the molecular taxonomic delineation of C. orientalis via universal DNA barcoding primers.The following new information regarding C. orientalis is provided in this study:Carnus orientalis is first recorded in Taiwan, filling the gap in its East Asian distribution. This is also the first record of Carnidae from Taiwan.Otus lettia (Hodgson, 1836) (Aves, Strigidae) is reported as a new host for C. orientalis, identified on a fallen fledgling.Co-amplification of the host's COI is reported in this study using the universal PCR primer set LCO1490/HCO2198. Additionally, the amplification of a COI-like pseudogene using a newly-designed primer set is detected through abnormal translated amino acid sequences and the occurrence of a stop codon.New specific primers for the COI gene of Carnus were designed in this study.The new distribution and ecological data of C. orientalis enhance our understanding of this species. The provision of new COI primers is anticipated to contribute to future studies employing DNA barcoding in bird-parasitic flies.

Thoracophelia Ehlers, 1897 is a genus of Opheliidae characterised by the body divided into three distinct regions, modified parapodia in chaetiger 10 and a ventral groove restricted to the posterior half of the body. To date, 18 species have been described in the genus. Amongst them, six species have been recorded in northeast Asia.A new species, Thoracophelia foliformis sp. nov., was discovered in the intertidal zone of the Yellow Sea, South Korea. This is the first Thoracophelia species report from the Yellow Sea. This new species is closely related to T. dillonensis (Hartman, 1938) from California and T. ezoensis Okuda, 1936 from Japan in having pectinate branchiae. However, the new species can be distinguished from the two species by the unique combination of the following characteristics: 15 pairs of wrinkled pectinate branchiae with 12–15 filaments at best development and a foliaceous mid-ventral plate in the pygidium instead of one or two thick ventral cirri. Detailed descriptions and illustrations of T. foliformis sp. nov. are provided. Sequences of the mitochondrial cytochrome c oxidase subunit I (COI), nuclear 18S ribosomal DNA (rDNA) and 28S rDNA of the new species were determined and analysed.

Abstract Background Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. Methods Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. Results The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28–30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74–18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. Conclusions This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided. Graphical abstract

DNA barcodes, short and unique DNA sequences, play a crucial role in sample identification when processing many samples simultaneously, which helps reduce experimental costs. Nevertheless, the low quality of long-read sequencing makes it difficult to identify barcodes accurately, which poses significant challenges for the design of barcodes for large numbers of samples in a single sequencing run. Here, we present a comprehensive study of the generation of barcodes and develop a tool, PRO, that can be used for selecting optimal barcode sets and demultiplexing. We formulate the barcode design problem as a combinatorial problem and prove that finding the optimal largest barcode set in a given DNA sequence space in which all sequences have the same length is theoretically NP-complete. For practical applications, we developed the novel method PRO by introducing the probability divergence between two DNA sequences to expand the capacity of barcode kits while ensuring demultiplexing accuracy. Specifically, the maximum size of the barcode kits designed by PRO is 2,292, which keeps the length of barcodes the same as that of the official ones used by Oxford Nanopore Technologies (ONT). We validated the performance of PRO on a simulated nanopore dataset with high error rates. The demultiplexing accuracy of PRO reached 98.29% for a barcode kit of size 2,922, 4.31% higher than that of Guppy, the official demultiplexing tool. When the size of the barcode kit generated by PRO is the same as the official size provided by ONT, both tools show superior and comparable demultiplexing accuracy.

INTRODUCTION. The creation of synthetic adeno-associated virus (AAV) vectors during gene therapy development is a labour-intensive and expensive process. The optimal solution to minimise the time and costs associated with gene therapy development lies in the improvement of methods aimed at assessing AAV vector biodistribution and transduction efficiency in vivo.AIM. This study aimed to develop a new bioinformatics-based assessment method for synthetic AAV vector libraries to analyse AAV vector biodistribution and transduction efficiency in vivo.MATERIALS AND METHODS. The production of synthetic AAV vectors involved assigning AAV serotype-specific barcodes (12-nucleotide tags flanked at the 5' end with a sequence encoding the green fluorescent reporter protein). Plasmids carrying unique barcodes were propagated in competent Escherichia coli XL10-Gold cells and used to create two AAV libraries: L1 with a viral genome count of 1010 and L2 with a viral genome count of 1011. AAV production involved HEK293T cell transfection. L1 and L2 library vectors were administered to C57Bl/6N mice by intravenous injection. DNA and RNA were isolated from transduced organs for analysis by next-generation sequencing. The obtained data on DNA and RNA barcode quantities in different murine organs were analysed to assess the biodistribution and transduction efficiency of synthetic AAVs. Barcodes were identified by aligning them to the expected sequences and counted. The resulting values were normalised to the quantity of barcodes in the original library.RESULTS. Seven viral constructs based on different AAV serotypes were created as part of two AAV libraries. Six of the AAV serotypes were synthetic (sAAV1, sAAV2, sAAV3, sAAV4, sAAV5, and sAAV6). Sequencing of murine organ samples revealed significant quantities of DNA barcodes from both AAV libraries in all organs except the brain. For the L1 library, RNA barcodes were detected at a sufficient level in 4 organs, including the skeletal muscles, the heart, the liver, and the adrenal glands. For the L2 library, in addition to the listed organs, sufficient RNA-barcode levels were observed in the gonads and the kidneys. According to transduction efficiency analysis based on RNA barcode levels adjusted for DNA barcodes, sAAV5 was considered the most promising variant for gene therapy of liver-related diseases, whereas sAAV2 and sAAV6 were recognised as holding the most promise for adrenal diseases.CONCLUSIONS. The developed bioinformatics-based assessment method for synthetic AAV vector libraries can analyse AAV vector biodistribution and transduction efficiency in the body. The presented approach has the potential for selecting optimal AAV vectors for specific organs and tissues in further gene therapy development.

Sterculia monosperma fruit is the dried and mature seed of Sterculia monosperma, which belongs to genus Sterculia. In China, this plant is a widely distributed arbor with a long planting history in Lingnan area. It is mainly cultivated in Guangdong, Guangxi, Fujian, Yunnan and Taiwan of China; it is also distributed in India, Vietnam and Indonesia, mostly cultivated artificially. S. monosperma fruit has many functions like warming the stomach and killing pests, but its name and characteristics are easily confused with other plants of genus Sterculia. However, the relevant research foundation is relatively weak. Pharmacognostical study can provide reference for its resource development and quality standard formulation. This study identifies the morphological and microscopic characteristics of S. monosperma fruit, as well as the microscopic identification of seed cross-section and powder. The DNA barcode sequences ITS2, psbA-trnH, matK and rbcL of S. monosperma were obtained through bidirectional sequencing, Kimura 2-Parameter (K2P) genetic distance was calculated, and the neighbor joining tree was established for clustering analysis. The results were as follows: (1) The morphological characteristics of S. monosperma fruit included dark red fruit shell and reddish brown or dark chestnut surface. It was hard in texture, with two thick yellowish endosperms inside. (2) The micro-morphological characteristics was that the exotesta was reddish brown, extremely thin, and brittle in texture; the mesotesta was black brown, thick, and hard in texture; the endotesta coat was light yellow and soft in texture. (3) The microscopic characteristics included the structure and arrangement of the exotesta stone cell, the grid cell structure of the mesotesta, the bead thickening of the cell wall of the endotesta cells, and the calcium oxalate cluster crystals. (4) Based on the ITS2 sequence, S. monosperma fruit could be effectively distinguished from other plants in genus Sterculia, while the matK sequence could effectively distinguish S. lanceolata from other plants in genus Sterculia. This study obtained the data on the morphological characteristics, micro-morphological characteristics and microscopic characteristics of S. monosperma fruit. Combined with the ITS2 barcode sequence, S. monosperma fruit can be effectively identified, which provides a scientific reference for the development of its germplasm resources and the formulation of relevant quality standards.

Abstract [Objective] The purpose of the research was to explore the evolutionary pattern of chloroplast genome in Elaeagnaceae, and to provide a basis for species identification and resource exploitation of Elaeagnaceae. [Methods] This study assembled four chloroplast genomes from Hippophae and Shepherdia, and compared the gene composition, repeats, and structural properties with the published Elaeagnaceae genomes. A phylogenetic tree was constructed based on sequences from these chloroplast genomes. The promising sites for DNA barcoding of Elaeagnaceae were identified by examining the highly divergent regions. [Results] It was found that the chloroplast genomes of each genus of Elaeagnaceae exhibited a significant degree of similarity in terms of tetrad structure and gene number and organization. Moreover, it was found that both IR regions and repeats of Hippophae and Shepherdia species tended to expand and increase more than those of Elaeagnus species. The phylogenetic tree based on chloroplast genomes from eighteen species of Elaeagnaceae showed that all Elaeagnus plants were grouped into a branch that exhibited an early differentiation, and all plants from Hippophae and Shepherdia were classified into another branch with a most recent common ancestry. Three potential DNA barcoding regions were examined with the ycf1 gene had the best effect. [Conclusion] This study revealed that the chloroplast genome structure of the Elaeagnus family had a conservative pattern. The IR regions and repeated sequence of the chloroplast genome had been extended and increased with the evolution of Elaeagnaceae. The selected DNA barcode sequences could distinguish the relationships among the genera of the family Elaeagnus and among the species within the genus Elaeagnus.

Homalernis fluctuosa Suzuki & Jinbo, sp. nov., is described and illustrated from Honshu, Shikoku, Kyushu, Tsushima Island, Amamiooshima Island, and Okinawajima Island, Japan. This is not only the first record of the genus Homalernis but also of the tribe Schoenotenini from warm temperate zones in the Palaearctic region. The association of males and females of the new species was confirmed based on the mitochondrial gene cytochrome oxidase submit 1 (COI). We discuss the taxonomic positions of two alleged Homalernis species from Malaysia and the taxonomic position of Homalernis within Schoenotenini.

The Pinna nobilis, a Mediterranean mollusc, has suffered population declines due to a massive mortality event associated with various factors including the parasite Haplosporidium pinnae. Some populations show resilience, possibly due to local environmental conditions. In this study, a molecular multiplex PCR method was developed using species‐specific primers targeting Internal Transcribed Spacer (ITS) regions of P. nobilis and P. rudis, allowing accurate species identification and hybrid detection. Samples from Mediterranean areas were analysed, including putative hybrids and individuals from five other bivalve species. DNA was isolated, ITS regions were amplified and sequenced, and phylogenetic analyses confirmed species differentiation and primer specificity. The multiplex‐PCR successfully identified P. nobilis, P. rudis, and their hybrids based on distinct amplicon patterns. This study highlights the value of molecular tools in species conservation, especially for monitoring and managing hybridization, supporting effective biodiversity conservation strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Alpine musk deer, red serow, and white-lipped deer coexist in the Nyenchen Tanglha Mountains of Tibet. We aimed to understand the mechanisms of their coexistence by studying their dietary preferences using DNA barcoding. All of the species exhibited broad dietary ranges with distinct food preferences. Furthermore, our findings revealed genus-level dietary specializations and the mechanisms facilitating their coexistence. The results of this study provide valuable insights for the development and implementation of effective conservation strategies and management measures in the local area. Wild ungulates play crucial roles in maintaining the structure and function of local ecosystems. The alpine musk deer (Moschus chrysogaste), white-lipped deer (Przewalskium albirostris), and red serow (Capricornis rubidus) are widely distributed throughout the Nyenchen Tanglha Mountains of Tibet. However, research on the mechanisms underlying their coexistence in the same habitat remains lacking. This study aimed to investigate the mechanisms underlying the coexistence of these species based on their dietary preferences through DNA barcoding using the fecal samples of these animals collected from the study area. These species consume a wide variety of food types. Alpine musk deer, white-lipped deer, and red serow consume plants belonging to 74 families and 114 genera, 62 families and 122 genera, and 63 families and 113 genera, respectively. Furthermore, significant differences were observed in the nutritional ecological niche among these species, primarily manifested in the differentiation of food types and selection of food at the genus level. Owing to differences in social behavior, body size, and habitat selection, these three species further expand their differentiation in resource selection, thereby making more efficient use of environmental resources. Our findings indicate these factors are the primary reasons for the stable coexistence of these species. [ABSTRACT FROM AUTHOR]

Taxonomy underpins biological research because names are needed for comparative analysis, conservation status, and public communication. Despite this, many species remain undescribed and are therefore vulnerable and unprotected, particularly in the tropics. Neotropical Staphylinidae beetles (Coleoptera) are among the insect groups most likely to contain significant unknown and/or cryptic diversity. Here we used an integrative taxonomic framework to conduct a preliminary review of one particularly diverse genus of Staphylinidae (Anotylus) in Área de Conservaciόn Guanacaste, northwestern Costa Rica. We began by DNA barcoding novel collections and using Barcode Index Numbers (BINs) as an estimate of taxonomic diversity; we found 18 provisional new species. We augmented this genetic analysis with a morphometric analysis of adult morphological characters and found that we could differentiate most provisional species by external morphology as well as by elevation of collection. All the most abundant species could be differentiated from each other by differences in body size. One BIN included slight (~1%) genetic variation that corresponded with some morphological differentiation suggesting the existence of two species within a DNA BIN. Our results support the efficacy of DNA barcoding collections of tropical insects as an effective biodiversity estimator, one that can be used as a primer for integrative taxonomic studies using BINs as species hypotheses. [ABSTRACT FROM AUTHOR]

Gastropods are ecologically significant and can be found in a wide range of habitats. Their taxonomy is complicated by their perplexing morphological characteristics. Therefore, a molecular based identification approach was used to confirm the variety of gastropod species commonly found on the coast during low tide at rocky shores. In the present investigation, nine species of gastropods were identified using the foresaid modern technique such as (Thalessa savignyi, Purpura persica, Turbo sparverius, Lunella coronata, Cellana karachiensis, Nerita albicilla, Nerita tristis, Ischnochiton australis, Astralium tentorriformis). T our knowledge among them three are news records (A. tentoriiformes, Ischnochiton australis and, Nerita tristis). The highest evolutionary divergence within the family of Nacellidae (0.917) followed by Turbinidae (0.169) and Neritidae (0.100) was noted. The results of this study will be helpful to taxonomists because meager attention was given to molecular- based identification from this region. [ABSTRACT FROM AUTHOR]

Nepal holds a high ichthyofaunal diversity, nevertheless, the molecular study of Nepalese fish is still in its early stages. The first record of a cyprinid fish species the Garra kempi Hora, 1921 for Nepal was reported from eastern Nepal's Lohandra River. Both morphology and molecular data affirmed the existence of G. kempi in Nepal's aquatic system. This species was previously reported from China (Tibet) and northern India. The maximum likelihood phylogenetic analysis and pairwise genetic distance based on Kimura 2 parameters using cytochrome oxidase subunit 1 (COI) segment sequences (665 bp) also confirmed the identity of the species as G. kempi. The COI sequences of specimens from Nepal formed a monophyletic clade with the sequence of G. kempi from northeastern India and the two contained a pairwise genetic distance of 1.8% only. The new record of G. kempi from Nepal warrants a detailed ichthyofaunal survey for documenting the fish diversity in Nepal. [ABSTRACT FROM AUTHOR]

The subfamily Molytinae (Coleoptera: Curculionidae), being the second largest group within the family Curculionidae, exhibits a diverse range of hosts and poses a serious threat to agricultural and forestry industries. We used 1,290 cytochrome c oxidase subunit I (COI) barcodes to assess the efficiency of COI barcodes in species differentiation and uncover cryptic species diversity within weevils of Molytinae. The average Kimura 2-parameter distances within species, genus, and subfamily were 2.90%, 11.0%, and 22.26%, respectively, indicating significant genetic differentiation at both levels. Moreover, there exists a considerable degree of overlap between intraspecific (0%–27.50%) and interspecific genetic distances (GDs; 0%–39.30%). The application of Automatic barcode gap discovery, Assemble Species by Automatic Partitioning, Barcode Index Number, Poisson Tree Processes (PTP), Bayesian Poisson Tree Processes (bPTP), and jMOTU resulted in the identification of 279, 275, 494, 322, 320, and 279 molecular operational taxonomic units, respectively. The integration of 6 methods successfully delimited species of Molytinae in 86.6% of all examined morphospecies, surpassing a threshold value of 3% GD (73.0%). A total of 28 morphospecies exhibiting significant intraspecific divergences were assigned to multiple MOTUs, respectively, suggesting the presence of cryptic diversity or population divergence. The identification of cryptic species within certain morphological species in this study necessitates further investigation through comprehensive taxonomic practices in the future. [ABSTRACT FROM AUTHOR]

Caridina boehmei is an ornamental shrimp native to Sulawesi that has been successfully cultured and marketed in Indonesia. However, molecular information about this shrimp is not yet available. This study aimed to characterize cytochrome c oxidase subunit I (CO1) and small subunit ribosomal RNA (18S rDNA) genes. Shrimp samples were extracted, and DNA was amplified using CO1 and 18S rDNA primers. The sequencing results were then analyzed by the neighbor-joining method. The 654 bp partial coding sequence (cds) of the CO1 and 380 bp partial cds of 18S rDNA have been successfully amplified from genomic DNA. The CO1 sequencing produced fragments consisting of 25.84% adenine, 20.95% cytosine, 19.11% guanine, and 34.10% thymine. The 18S rDNA sequencing produced fragments consisting of 24.47% adenine, 26.05% cytosine, 28.16% guanine, and 21.32% thymine. Phylogenetic analysis based on the CO1 gene revealed that C. boehmei was grouped with C. variabilis (MK190014), C. brachydactyla (MK190011), and Caridina sp. (MK190012). Phylogenetic analysis based on the 18S rDNA gene revealed that C. boehmei was identical to Caridina sp. Suphan Buri-1 MF622000, C. serratirostris KP725708, C. multidentata JF346236, C. formosae GQ131924, and Caridina sp. Suphan Buri-2 MF622001. The lowest-highest pairwise distance based on CO1 and 18S rDNA was consecutively 0.0017-0.2247 (0.17-22.47%) and 0.0000-0.1218 (0-12.18%). [ABSTRACT FROM AUTHOR]

The Leptomias group represents one of the most diverse taxonomic group of weevils in the Qinghai‐Tibet Plateau and its adjacent areas. Despite the potential of hidden diversity, relatively few comprehensive studies have been conducted on species diversity in this taxonomic group. In this study, we performed DNA barcoding analysis for species of the Leptomias group using a comprehensive DNA barcode dataset that included 476 sequences representing 54 morphospecies. Within the dataset, our laboratory contributed 474 sequences, and 390 sequences were newly generated for this study. The average Kimura 2‐parameter distances among morphospecies and genera were 0.76% and 19.15%, respectively. In 94.4% of the species, the minimum interspecific distances exceeded the maximum intraspecific distances, indicating the presence of barcode gaps in most species of Leptomias group. The application of Automatic Barcode Gap Discovery, Assemble Species by Automatic Partitioning, Barcode Index Number, Bayesian Poisson tree processes, jMOTU, and Neighbor‐joining tree methods revealed 45, 45, 63, 54, and 55 distinct clusters representing single species, respectively. Additionally, a total of four morphospecies, Leptomias kangmarensis, L. midlineatus, L. siahus, and L. sp.9RL, were found to be assigned to multiple subclade each, indicating the geographical divergences and the presence of cryptic diversity. Our findings of this study demonstrate that Qinghai‐Tibet Plateau exhibits a higher species diversity of the Leptomias group, and it is imperative to investigate cryptic species within certain morphospecies using integrative taxonomic approaches in future studies. Moreover, the construction of a DNA barcode reference library presented herein establishes a robust foundational dataset to support forthcoming research on weevil taxonomy, phylogenetics, ecology, and evolution. [ABSTRACT FROM AUTHOR]

Simple Summary: In the species-diverse butterfly genus Polyommatus, speciation is often driven by rapid changes in chromosome number and structure, resulting in cryptic species. These cryptic species are morphologically similar, but usually can be recognized relatively easily using chromosomal markers. In this work, we show that similar chromosome numbers can independently evolve in different phylogenetic lineages, resulting in species that are difficult to distinguish using routine cytogenetic techniques. We also demonstrate that the combined analysis of chromosomes and mitochondrial DNA barcodes is a simple and effective tool for identifying such cryptic species. The detection of cryptic species in complexes that have undergone recent speciation is often difficult, since many standard nuclear markers have not yet accumulated differences between closely related taxa, and differences in mitochondrial markers can be leveled out due to mitochondrial introgressions. In these cases, the use of derived chromosomal characters such as non-ancestral chromosomal numbers and/or unusual karyotype features may be a solution to the species delimitation problem. However, non-ancestral but similar karyotypes may arise secondarily as a result of homoplastic evolution, and their interpretation as homologies may lead to incorrect taxonomic conclusions. In our study, we show that the combined use of mitochondrial DNA barcodes and karyotypes helps to solve this problem and identifies cryptic species in situations where each of these markers does not work individually. Using this approach, we show that the fauna of Armenia and adjacent Iran includes the following cryptic taxa of the Polyommatus ripartii species complex (haploid chromosome number, n in parentheses): P. ripartii paralcestis (n = 90), P. ripartii kalashiani, subsp. nov (n close to 90), P. emmeli, sp. nov. (n = 77–79), P. keleybaricus, sp. nov. (n = 86), P. demavendi belovi (n = 73–75), P. demavendi antonius, subsp. nov. (n = 71–73), P. admetus anatoliensis (n = 79) and P. eriwanensis (n = 29–34). Polyommatus admetus yeranyani is synonymized with P. admetus anatoliensis. [ABSTRACT FROM AUTHOR]

Hypostomus is the most species-rich genus within the family Loricariidae and is widely distributed throughout the Neotropical region. Nonetheless, the diversity and distribution of these species have still large knowledge gaps. This scenario is more significant in some regions, such as the northeast of Brazil. In this region, the first species of the genus, H. pusarum , was described in the Northeast Caatinga and Costal Drainages ecoregion. Six congeners were subsequently described in the same ecoregion, all sharing the same colour pattern making them difficult to distinguish. All of them are collectively referred to as the H. pusarum complex. The present work seeks to clarify the diversity that constitutes the H. pusarum complex through an integrative study using molecular and morphological data. The results indicate that H. carvalhoi , H. jaguribensis , H. nudiventris , H. papariae , and H. salgadae are all junior synonyms of H. pusarum. However, one of the morphotypes that occurs in the Parnaíba River is a new species that differs from the others by the absence of a developed medial buccal papilla and the presence of a pre-anal plate. The data provided here highlight the importance of integrative taxonomy for assessing diversity in complex and diverse groups in the Neotropics. [ABSTRACT FROM AUTHOR]

Current knowledge of the global diversity of schizophoran flies, the megadiverse group that evolved during the episode of rapid dipteran radiations in the Early Cenozoic, is far from satisfactory. A significant part of this radiation is flesh flies (Sarcophagidae), which include about 3000 known species. Miltogramma Meigen, the largest genus of the flesh fly subfamily Miltogramminae, includes 119 known species with a peak of diversity in the Middle East and Central Asia. The taxonomy of the genus is acknowledged to be challenging, partly due to logistical issues associated with the deposition of type specimens in less accessible Russian institutions. Six new distinctive species of Miltogramma collected during recent expeditions to Iran and Uzbekistan are described using methods of integrative taxonomy: Miltogramma manouchehrii Szpila & Pape, sp. nov. , M. mikolajczyki Szpila & Pape, sp. nov. , M. rognesi Szpila & Pape, sp. nov. , M. soszynskii Szpila & Pape, sp. nov. , M. thompsoni Szpila & Pape, sp. nov. , and M. vervesi Szpila & Pape, sp. nov. Flies are characterized by secondary sexual dimorphism expressed in the form of species-specific chaetotaxy of the male fore tarsus, a character related to courtship behaviour. The reconnaissance surveys to both of the mentioned countries performed in the last decade brought information about a large number of potential new species of Miltogramma , despite the taxonomic work performed in this area in the past. Describing the majority of them requires extensive revisionary work, which involves accessing the type specimens and applying modern methods of species delimitation. [ABSTRACT FROM AUTHOR]

The genus Chamaecyparis comprises seven species (C. formosensis, C. hodginsii, C. lawsoniana, C. nootkatensis (Callitropsis nootkatensis), C. obtusa, C. pisifera, and C. thyoides). Accurate species identification is necessary for proper use and economic value of wood. Species identification of woods is generally based on anatomical analysis; however, C. obtusa and C. pisifera wood have similar microscopic morphology, which makes species identification impossible. Therefore, the molecular identification of species in wood of the genus Chamaecyparis is required. In this study, six candidate DNA barcode genes (trnP-GGG, ycf1b, clpP, accD, ycf2, and rps16) in the chloroplast of Chamaecyparis were identified with nucleotide diversity values higher than the arbitrary value of 0.02. Each gene was evaluated for species identification using phylogenetic analysis by genes registered at NCBI (42 sequences each for trnP-GGG, ycf1b, clpP, accD, and ycf2, and 50 sequences for rps16). The genes trnP-GGG, clpP, and rps16 could not be distinguished between C. pisifera and C. formosensis. However, ycf1b, accD, and ycf2 could be distinguished between all Chamaecyparis species. These results suggest the use of the chloroplast genes ycf1b, accD, and ycf2 as DNA barcodes for species identification in Chamaecyparis, including C. obtusa and C. pisifera, based on the reported genetic information to date. [ABSTRACT FROM AUTHOR]

Nepal holds a high ichthyofaunal diversity, nevertheless, the molecular study of Nepalese fish is still in its early stages. The first record of a cyprinid fish species the Garra kempi Hora, 1921 for Nepal was reported from eastern Nepal's Lohandra River. Both morphology and molecular data affirmed the existence of G. kempi in Nepal's aquatic system. This species was previously reported from China (Tibet) and northern India. The maximum likelihood phylogenetic analysis and pairwise genetic distance based on Kimura 2 parameters using cytochrome oxidase subunit 1 (COI) segment sequences (665 bp) also confirmed the identity of the species as G. kempi. The COI sequences of specimens from Nepal formed a monophyletic clade with the sequence of G. kempi from northeastern India and the two contained a pairwise genetic distance of 1.8% only. The new record of G. kempi from Nepal warrants a detailed ichthyofaunal survey for documenting the fish diversity in Nepal. [ABSTRACT FROM AUTHOR]

Four species of spiders intercepted at domestic ports were studied in this paper. The CO Ⅰsequences of which were amplified by PCR and compared with the CO Ⅰsequences of five species recorded in GenBank. And the sequence characteristics were analyzed and the phylogenetic tree was constructed. The results showed that there were significant genetic differences among different species of Latrodectus, and the CO Ⅰsequence could be used as a DNA barcode for molecular identification of different species of Latrodectus. [ABSTRACT FROM AUTHOR]

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

Sterculia monosperma fruit is the dried and mature seed of Sterculia monosperma, which belongs to genus Sterculia. In China, this plant is a widely distributed arbor with a long planting history in Lingnan area. It is mainly cultivated in Guangdong, Guangxi, Fujian, Yunnan and Taiwan of China; it is also distributed in India, Vietnam and Indonesia, mostly cultivated artificially. S. monosperma fruit has many functions like warming the stomach and killing pests, but its name and characteristics are easily confused with other plants of genus Sterculia. However, the relevant research foundation is relatively weak. Pharmacognostical study can provide reference for its resource development and quality standard formulation. This study identifies the morphological and microscopic characteristics of S. monosperma fruit, as well as the microscopic identification of seed cross-section and powder. The DNA barcode sequences ITS2, psbA-trnH, matK and rbcL of S. monosperma were obtained through bidirectional sequencing, Kimura 2-Parameter(K2P)genetic distance was calculated, and the neighbor joining tree was established for clustering analysis. The results were as follows:(1)The morphological characteristics of S. monosperma fruit included dark red fruit shell and reddish brown or dark chestnut surface. It was hard in texture, with two thick yellowish endosperms inside.(2)The micro-morphological characteristics was that the exotesta was reddish brown, extremely thin, and brittle in texture; the mesotesta was black brown, thick, and hard in texture; the endotesta coat was light yellow and soft in texture.(3)The microscopic characteristics included the structure and arrangement of the exotesta stone cell, the grid cell structure of the mesotesta, the bead thickening of the cell wall of the endotesta cells, and the calcium oxalate cluster crystals.(4)Based on the ITS2 sequence, S. monosperma fruit could be effectively distinguished from other plants in genus Sterculia, while the matK sequence could effectively distinguish S. lanceolata from other plants in genus Sterculia. This study obtained the data on the morphological characteristics, micro-morphological characteristics and microscopic characteristics of S. monosperma fruit. Combined with the ITS2 barcode sequence, S. monosperma fruit can be effectively identified, which provides a scientific reference for the development of its germplasm resources and the formulation of relevant quality standards. [ABSTRACT FROM AUTHOR]

Genetic barcodes and microsatellites are some of the identification methods in taxonomy and biodiversity research. It is important to establish a relationship between microsatellite quantification and genetic information in barcodes. In order to clarify the association between the genetic information in barcodes (expressed as Shannon’s Measure of Information, SMI) and microsatellites count, a total of 330,809 DNA barcodes from the BOLD database (Barcode of Life Data System) were analyzed. A parallel sliding-window algorithm was developed to compute the Shannon entropy of the barcodes, and this was compared with the quantification of microsatellites like (AT)n, (AC)n, and (AG)n. The microsatellite search method utilized an algorithm developed in the Java programming language, which systematically examined the genetic barcodes from an angiosperm database. For this purpose, a computational tool named Mfind was developed, and its search methodology is detailed. This comprehensive study revealed a broad overview of microsatellites within barcodes, unveiling an inverse correlation between the sumz of microsatellites count and barcodes information. The utilization of the Mfind tool demonstrated that the presence of microsatellites impacts the barcode information when considering entropy as a metric. This effect might be attributed to the concise length of DNA barcodes and the repetitive nature of microsatellites, resulting in a direct influence on the entropy of the barcodes. [ABSTRACT FROM AUTHOR]

The predaceous water beetle Menthles indicus is reported after a lapse of about five decades from India. The molecular phylogeny of the species has been described using CO1 gene sequence, which shows the species closely related to Methles cribratellus, by a genetic distance of 8–9%. [ABSTRACT FROM AUTHOR]

The effectiveness of plant-based therapies depends on precise botanical identification. However, tribal communities and practitioners sometimes misclassify medicinal plants due to a lack of scientific understanding. Plants cannot always be identified simply by morphology; the DNA-based approach uses a small segment of genomic DNA to authenticate medicinal plant species. Many tribal communities in Bangladesh have utilized plants for healing purposes for centuries. Therefore, this research used DNA sequences to identify Bangladeshi ethnomedicinal plants. DNA was extracted from the fresh leaves of eight important medicinal plants using the AddPrep Genomic DNA Extraction Kit and stored at − 20 °C. Three important loci (matK, rbcL and psbA-trnH) were amplified using PCR, and the sequences of these regions were analyzed for identification. rbcL and matK showed amplification success rates of 100% and 75% respectively, while for psbA-trnH the level was 50%, which subsequently led to the production of a total of 18 novel sequences. To validate the taxonomic identity of the samples, a MEGABLAST sequence comparison was performed, with nine species-level (50%) and nine genus-level (50%) identifications confirmed. The neighbor-joining method was used to construct four phylogenetic trees. The sequences obtained from the plant (leaf) samples and downloaded sequences from NCBI were employed, with the topologies of the trees being identical. Our findings demonstrate that partial genome sequencing is a viable way of identifying medicinal plants and closely related species utilizing the three plastid loci rbcL, matK and psbA-trnH, which are efficient and feasible DNA markers for plant authenticity. [ABSTRACT FROM AUTHOR]

The evergreen tree species Aquilaria sinensis holds significant economic importance due to its specific medicinal values and increasing market demand. However, the unrestricted illegal exploitation of its wild population poses a threat to its survival. This study aims to contribute to the conservation efforts of A. sinensis by constructing a library database of DNA barcodes, including two chloroplast genes (psbA-trnH and matK) and two nuclear genes (ITS and ITS2). Additionally, the genetic diversity and structure were estimated using inter-simple sequence repeats (ISSR) markers. Four barcodes of 57 collections gained 194 sequences, and 1371 polymorphic bands (98.63%) were observed using DNA ISSR fingerprinting. The Nei's gene diversity (H) of A. sinensis at the species level is 0.2132, while the Shannon information index (I) is 0.3128. The analysis of molecular variance revealed a large significant proportion of total genetic variations and differentiation among populations (Gst = 0.4219), despite a relatively gene flow (Nm = 0.6853) among populations, which were divided into two groups by cluster analysis. There was a close genetic relationship among populations with distances of 0.0845 to 0.5555. This study provides evidence of the efficacy and dependability of establishing a DNA barcode database and using ISSR markers to assess the extent of genetic diversity A. sinensis. Preserving the genetic resources through the conservation of existing populations offers a valuable proposition. The effective utilization of these resources will be further deliberated in subsequent breeding endeavors, with the potential to breed agarwood commercial lines. [ABSTRACT FROM AUTHOR]

Species of the cleptoparasitic bee genus Triepeolus Robertson, 1901 (Hymenoptera: Apidae: Nomadinae) of which the female has a trapezoidal or triangular pseudopygidial area with bright, reflective setae and a concave apical margin are revised. This entirely New World group includes the widely known species T. simplex Robertson, 1903 and is thus termed the simplex species group. A total of 18 species in the T. simplex group are recognized as valid, of which seven are newly named and described—T. apache sp. nov., T. eumeniformis sp. nov., T. hirsutus sp. nov., T. oblongimacula sp. nov., T. parahirsutus sp. nov., T. paucipunctatus sp. nov., and T. shoshone sp. nov., all from North America. Eleven redundant names are newly synonymized under three valid ones as follows: Epeolus rugulosus Cockerell, 1917 syn. nov., E. metatarsalis Friese, 1921 syn. nov., and T. bilunatus Cockerell, 1949 syn. nov. under Triepeolus mexicanus (Cresson, 1878); E. lectiformis Cockerell, 1925 syn. nov., T. lusor Cockerell, 1925 syn. nov., and T. junctus Mitchell, 1962 syn. nov. under T. rhododontus Cockerell, 1921; and E. permixtus Cockerell, 1923 syn. nov., T. brunnescens Cockerell & Sandhouse, 1924 syn. nov., T. pacis Cockerell, 1925 syn. nov., E. sarothrinus Cockerell, 1929 syn. nov., and E. sarothrinus var. confluens Cockerell, 1929 syn. nov. under T. segregatus (Cockerell, 1900). Species limits were established using an integrative systematics approach, namely considering morphological and biogeographic evidence as well as DNA barcode data. Taxon concepts are revised for all species in the T. simplex group, with morphological diagnoses and keys presented to enable their identification. Known information on the ranges and ecology of the treated species is summarized.

Trypanosomes are blood parasites capable of infecting nearly any vertebrate. Many Neotropical primates frequently host trypanosomes and are considered potential reservoirs for Trypanosoma cruzi and other human-pathogenic trypanosomatids. However, diagnostic methods originally developed for detecting these trypanosomatids in humans and domestic species must be validated to reliably diagnose infections in non-human primates. Without such validation, taxonomic biases and incorrect assignments of wildlife reservoirs can occur. The white-footed tamarin (Oedipomidas leucopus), a primate endemic to northwestern Colombia, is classified by the World Health Organization as a reservoir of T. cruzi. However, this classification is based on studies with small sample sizes, ambiguous diagnostic methods, and questionable geographic records. In this study, the 18S ribosomal RNA gene was amplified via PCR and sequenced to estimate trypanosome infection rates and identify species in natural populations of O. leucopus across a wide geographic range, as well as in (ex situ) specimens. This molecular approach was also compared with traditional microscopy diagnosis using blood smears. The molecular diagnosis revealed that over 60% of the tested specimens were infected, whereas traditional microscopy resulted in 58% false negatives compared to the molecular method. A Bayesian phylogeny of the 18S gene identified T. minasense as the sole trypanosomatid species present in O. leucopus, with no detections of T. cruzi or other trypanosomatids of concern to human or domestic animal health. This study highlights the risk of overestimating the presence of human-infecting trypanosomes, such as T. cruzi, in tamarins and other vertebrates, and underscores the importance of validating diagnostic methods to accurately assess the zoonotic potential of wild species. Accurate identification of wildlife reservoirs is essential for understanding parasite life cycles and implementing effective management and conservation strategies for primates and other potential reservoirs.

A new species of the genus Acrossus Mulsant, 1842, Acrossus baei sp. nov. from South Korea, is described and illustrated on the basis of morphology and mitochondrial COI sequences. The species was compared with four related species; Acrossus atratus (Waterhouse, 1875), A. humerospinosus (Petrovitz, 1958), A. luridus (Fabricius, 1775), and A. superatratus (Nomura & Nakane, 1951). The taxonomic status and diagnostic characters of the new species are discussed. A key to species of the genus Acrossus in the Korean Peninsula is given. [ABSTRACT FROM AUTHOR]

To update the record of mosquito diversity in the state of Morelos, Mexico, mosquito collections were conducted in both physiographical regions of Morelos: the Neo-volcanic Axis and the South Mountain Range. Immature stages were collected from aquatic habitats, while adult mosquitoes were collected using CDC light traps, Shannon traps and approaching/landing on the collecting personnel. All specimens were identified using taxonomic keys and the mitochondrial cytochrome C oxidase subunit 1 (COI). Following Mexico State, this is the second study of mosquito taxonomy of a complete Mexican state using COI barcoding. In Morelos, a total of 58 species of mosquito occurs, of which we report 12 for first time (Aedes vexans, Ae. euplocamus, Ae. vargasi, Psorophora cilipes, Ps. lineata, Culex restrictor, Cx. interrogator, Cx. nigripalpus, Cx. tarsalis, Cx. sandrae, Cx. lactator and Toxorhynchites moctezuma). Morelos now ranks sixth in mosquito diversity among Mexican states. Phylogenetic trees were constructed for 32 species, including Ae. shannoni, which is a new record for GenBank databases. Additionally, we report COI barcode sequences of Culiseta dugesi Dyar and Knab, which is resurrected from the synonymy of Cs. particeps (Adams). Fourth-instar larvae and adult females of Cs. dugesi were redescribed. This brings the number of mosquito species in Mexico to 248. Ecological and distributional notes for the new records, the medical importance of mosquito species in Morelos, and an identification key for larvae and females of the known Culiseta species occurring in Mexico are also included. [ABSTRACT FROM AUTHOR]

Synanthropic silverfish are the best-known and most widely distributed insects of the order Zygentoma. However, there is a great gap in the knowledge and confusion about the geographic distribution and the diagnostic characteristics that allow their identification. In this work, we provide an exhaustive and deep analysis of the most common 9 synanthropic silverfish of the world, combining previously published and newly derived morphological and molecular data. Updated descriptions of Ctenolepisma calvum (Ritter, 1910) and Ctenolepisma (Sceletolepisma) villosum (Fabricius, 1775) are included, and morphological remarks, illustrations, and photographs of the remaining synanthropic species are provided to clarify their diagnosis and differentiation among them and from other free-living species. In addition, Ctenolepisma targionii (Grassi and Rovelli, 1889) is synonymized with C. villosum. A molecular phylogeny is presented based on the COI sequences of all the synanthropic species deposited in BOLD and GenBank, with 15 new sequences provided by this study. This has allowed us to detect and correct a series of identification errors based on the lack of morphological knowledge of several species. Moreover, 2 different lineages of Ctenolepisma longicaudatum Escherich, 1905 have also been detected. To help future studies, we also provide a taxonomic interpretation guide for the most important diagnostic characters of the order Zygentoma, as well as an identification key for all the Synanthropic studied species. Finally, an approximation of the global distribution of synanthropic silverfish is discussed. Several new records indicate that the expansion of these species, generally associated with the transport of goods and people, is still far from over. [ABSTRACT FROM AUTHOR]