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Ectomycorrhizal (ECM) fungi are important tree symbionts within forests. The biogeography of ECM fungi remains to be investigated because it is challenging to observe and identify species. Because most ECM plant taxa have a Holarctic distribution, it is difficult to evaluate the extent to which host preference restricts the global distribution of ECM fungi. To address this issue, we aimed to assess whether host preference enhances the endemism of ECM fungi that inhabit dipterocarp rainforests. Highly similar sequences of 175 operational taxonomic units (OTUs) for ECM fungi that were obtained from Lambir Hill's National Park, Sarawak, Malaysia, were searched for in a nucleotide sequence database. Using a two‐step binomial model, the probability of presence for the query OTUs and the registration rate of barcode sequences in each country were simultaneously estimated. The results revealed that the probability of presence in the respective countries increased with increasing species richness of Dipterocarpaceae and decreasing geographical distance from the study site (i.e. Lambir). Furthermore, most of the ECM fungi were shown to be endemic to Malaysia and neighbouring countries. These findings suggest that not only dispersal limitation but also host preference are responsible for the high endemism of ECM fungi in dipterocarp rainforests. Moreover, host preference likely determines the areas where ECM fungi potentially expand and dispersal limitation creates distance–decay patterns within suitable habitats. Although host preference has received less attention than dispersal limitation, our findings support that host preference has a profound influence on the global distribution of ECM fungi. [ABSTRACT FROM AUTHOR]

For two decades, DNA barcoding and, more recently, DNA metabarcoding have been used for molecular species identification and estimating biodiversity. Despite their growing use, few studies have systematically evaluated these methods. This study aims to evaluate the efficacy of barcoding methods in identifying species and estimating biodiversity, by assessing their consistency with traditional morphological identification and evaluating how assignment consistency is influenced by taxonomic group, sequence similarity thresholds and geographic distance. We first analysed 951 insect specimens across three taxonomic groups: butterflies, bumblebees and parasitic wasps, using both morphological taxonomy and single‐specimen COI DNA barcoding. An additional 25,047 butterfly specimens were identified by COI DNA metabarcoding. Finally, we performed a systematic review of 99 studies to assess average consistency between insect species identity assigned via morphology and COI barcoding and to examine the distribution of research effort. Species assignment consistency was influenced by taxonomic group, sequence similarity thresholds and geographic distance. An average assignment consistency of 49% was found across taxonomic groups, with parasitic wasps displaying lower consistency due to taxonomic impediment. The number of missing matches doubled with a 100% sequence similarity threshold and COI intraspecific variation increased with geographic distance. Metabarcoding results aligned well with morphological biodiversity estimates and a strong positive correlation between sequence reads and species abundance was found. The systematic review revealed an 89% average consistency and also indicated taxonomic and geographic biases in research effort. Together, our findings demonstrate that while problems persist, barcoding approaches offer robust alternatives to traditional taxonomy for biodiversity assessment. [ABSTRACT FROM AUTHOR]

DNA barcodes (i.e., gene sequences used as identifiers for species identification) constitute a very useful tool in invasive pest research. Using them, we studied, for the first time, the origin and population structure of the alien Cydia pomonella in Africa, a major apple pest. We sequenced a fragment of the mitochondrial gene Cytochrome c oxidase subunit I (COI) from C. pomonella caterpillars collected in northern Algeria and compared them with DNA barcodes from the rest of the world available at public repositories. The phylogeny built upon that COI database supports the European origin of the species: haplotypes at the base of the phylogeny were registered in this continent. In Algeria, the results suggest that this pest could have been introduced from Europe, with two unique African mutations probably favoured by C. pomonella multivoltinism and large population sizes. Population genetic analyses at three Algerian localities showed neither structure nor founder effects. However, to confirm the underlying process of the observed population patterns, it is necessary to perform analyses using genes with higher mutation rates. Compared to higher resolution markers, mitochondrial DNA barcodes are still a cost‐effective tool for taxonomic determination of unknown specimens and, sometimes, may also allow a preliminary tracing of its origin. This is crucial to detect and stop the spread of invasive pests (i.e., borders). We encourage further studies including functional genes to assess whether any mutations are promoting C. pomonella adaptation to the Northern Africa environment. [ABSTRACT FROM AUTHOR]

Sand flies (Diptera: Psychodidae: Phlebotominae) are blood‐feeding insects that transmit the protozoan parasites Leishmania spp. and various arboviruses. The Balkan region, including the Republic of Kosovo, harbours a diverse sand fly fauna. Vector species of Leishmania infantum as well as phleboviruses are endemic; however, recent data are scarce. We performed a cross‐sectional study to update the current sand fly distribution in Kosovo and assess biological as well as environmental factors associated with sand fly presence. CDC light trapping was conducted at 46 locations in 2022 and 2023, specifically targeting understudied regions in Kosovo. Individual morphological species identification was supported by molecular barcoding. The occurrence data of sand flies was used to create distribution maps and perform environmental analyses, taking elevation, wind speed and climate‐related factors into account. In addition, PCR‐based blood meal analysis and pathogen screening were conducted. Overall, 303 specimens of six sand fly species were trapped, predominated by Phlebotomus neglectus (97%). Barcodes from eight of nine known endemic sand fly species were obtained. Combining our data with previous surveys, we mapped the currently known sand fly distribution based on more than 4000 specimens at 177 data points, identifying Ph. neglectus and Ph. perfiliewi as the predominant species. Environmental analyses depicted two geographical groups of sand flies in Kosovo, with notable differences between the species. In total, 223 blood meals of five sand fly species were analysed. Of seven identified host species, the predominant blood meal source was observed to be cattle, but the DNA of dogs and humans, among others, was also detected. This study assessed biological as well as ecological factors of sand fly occurrence, which should help better understand and evaluate potential hot spots of disease transmission in Kosovo. [ABSTRACT FROM AUTHOR]

The control authorities of timber trade lack quick and easy methods to verify the legality of the traded product. This includes the clear identification of the timber species. In this study, a simple genetic method, the macroarray technique, was modified and optimised for this purpose. The internal transcribed spacer (ITS) was used as barcode region. Specific probes were designed and tested using this barcode of trade-relevant, CITES protected wood species and their look-alikes. The results reveal that 61 of these probes are suitable for identification purposes. Hence, 32 of the 50 investigated species can be clearly identified using the developed macroarray. Additional 15 species are represented by probes in groups of two to three species, which enables a delimitation of the species present. For four species, no identification was possible applying the probes created. The findings indicate the potential of time and cost savings compared to currently established methods in species control in the timber trade. [ABSTRACT FROM AUTHOR]

Coprinellus disseminatus and other morphologically similar species are widely dispersed worldwide and are commonly referred to as "fairy inkcap". Based on the molecular phylogenetic study and morphological observation, a thorough investigation was carried out utilising 74 collections of related species that were gathered from seventeen provinces and five Chinese fungaria between 1998 and 2023 and revealed 11 lineages of "fairy inkcap", nine of which were found in China, and which belonged to the two genera Coprinellus and Tulosesus. In sect. Disseminati, genetic diversities (π), and fixation index (Fst) amongst lineages were computed, and a haplotype-based network was established to ascertain the relationships amongst each clade. A new section of Coprinellus, sect. Aureodisseminati, were discovered. In addition, four new species (C. aureodisseminatus, C. austrodisseminatus, C. parcus, and C. velutipes), a new subspecies of C. disseminatus, a new combination (Tulosesus pseudodisseminatus), the first discovery of epigamous type of C. magnoliae and a new record to China (T. subdisseminatus) were also identified and thoroughly described with accompanying illustrations. Their differences in macro- and micro-features, as well as their character sequence, were discussed. [ABSTRACT FROM AUTHOR]

• South Africa ranks third in global flowering plant barcode data. • The country has submitted more than 12,000 DNA barcode records to BOLD. • Endemic families and taxa are underrepresented, leaving significant data gaps. • Underutilised resources and funding constraints hinder data growth in the region. • Urgent need for collaboration, targeted sampling, and database harmonisation. South Africa, a global biodiversity hotspot, faces escalating threats to its rich plant diversity, including habitat loss, climate change, invasive species, and illegal harvesting. These threats are further exacerbated by the country's taxonomic impediment, which hinders both conservation and sustainable development efforts. This paper assesses the efficiency of DNA barcoding as a tool for species identification and biodiversity conservation within the South African context. While DNA barcoding offers promising applications in conservation, significant gaps and challenges persist. We provide a comprehensive overview of plant barcoding initiatives in South Africa – by querying the public data portal for plant barcoding records, 12,456 published specimen records encompassing 159 families and ca. 3,449 species were returned. These numbers highlight historical progress, database contributions, technological advancements, and taxonomic coverage. Despite South Africa contributing the third-highest number of Magnoliophyta records to the Barcode of Life Data System (BOLD), significant gaps in endemic families and geographic regions highlight the urgent need for targeted DNA barcoding initiatives and increased collaboration, as only ca. 16 % of the known flora has been barcoded. The underutilisation of BOLD and financial constraints pose significant barriers to expanding plant barcoding records. However, advancements in sequencing technologies offer cost-effective solutions. We advocate for concerted efforts to enhance DNA barcoding utilisation, harmonise databases, and prioritise sampling of underrepresented taxa to effectively preserve South Africa's diverse plant life. [ABSTRACT FROM AUTHOR]

Overfishing and unsustainable practices have caused drastic declines in shark populations worldwide; these decreases are largely attributed to the demand for shark products (e.g., fins and meat) and shark bycatch associated with the global fishing industry. In an effort to understand the species composition of the shark fin trade in Singapore – a globally significant trade hub, we collected and genetically identify a total of 6840 shark fins collected between January 2021 and February 2022. We then adopted a Bayesian modelling approach to understand how the identified species contributed to the overall trade within Singapore. The Singapore market appears distinct in terms of species composition when compared to the markets of Hong Kong and mainland China. In Singapore 81% of samples belonged to ten species, whereas, in Hong Kong and mainland China a single species dominated the trade. Of those identified in Singapore, the IUCN considers six to be threatened species and eight are listed in CITES Appendix II (in online). The differences in species composition between East and Southeast Asian markets suggest that different supply chains are sustaining these hubs. Given these differences, it is important that conservation, sustainable fisheries management and policy decisions are based upon the findings from multiple trade centres. [ABSTRACT FROM AUTHOR]

This work demonstrates for the first time using molecular markers alongside chromosomes and the intermediate phenotype that a specimen of Dorcadion is an accidental first‐generation hybrid. The analyses were based on two mitochondrial and five nuclear molecular markers. While analysing the putative parental taxa of the hybrid, it was revealed that even Dorcadion lugubre lugubre Kraatz is likely a hybridogenic taxon. Coincidentally, a new lead was discovered concerning the parental taxa of Dorcadion aethiops (Scopoli), already known as a hybridogenic species. This new evidence of hybridisation adds to the recent publications to strengthen the hypothesis that, in Dorcadionini, this phenomenon is relatively frequent, and speciation is often reticulate: New taxa can appear through introgressive hybridisation, explaining in part the extreme species richness of the group. [ABSTRACT FROM AUTHOR]

Georgia is part of two biodiversity hotspots, the Caucasus and the Irano-Anatolian, both of which are characterized by high biological diversity and endemism rates. Eighty-one species of small mammals (Eulipotyphla, Chiroptera, Rodentia, and Leporidae) have been recorded in Georgia so far based on morphological identification. However, a comprehensive understanding of small mammal diversity requires integrative taxonomic approaches due to the cryptic nature and sympatric distribution of some taxa. Here, to develop a DNA barcode library, we re-evaluated the small mammal diversity of Georgia. Samples for DNA studies were collected during the last nine years throughout Georgia. The Cytochrome Oxidase Subunit I (COI) region of the mitochondrial genome was barcoded for collected samples, and a DNA barcode library was assembled. As a result, nearly 80% of the known species of Georgian small mammals were covered. Two species, Microtus rossiaemeridionalis (Ognev, 1924) and Myotis tschuliensis (Kuzyakin, 1935), were detected for the first time in Georgia. Furthermore, several problems were identified within the Chiroptera and Rodentia orders, requiring further studies. [ABSTRACT FROM AUTHOR]

Caloptilia roscipennella (Hübner, 1796) is a leaf‐mining moth native to Central and Southern Europe. In this publication, we provide the first report of the occurrence of Caloptilia roscipennella in India. The identification was performed using an integrative approach, combining molecular (barcoding, COI) and morphological analysis (forewing pattern). In 2020, the larvae were initially found scraping in the silvery‐white mine and later feeding in folded or rolled leaves of the common walnut Juglans regia L. In the subsequent years, the species was found only in the same places. Two generations were recorded per year (May–June and August–September) and leaf miners overwintered as adults and reappeared in the early spring. Damage ranging from 15% to 20% was observed on the leaves of J. regia. Further efforts related to C. roscipennella should concentrate on evaluating its geographic distribution and its impact on walnut trees. Several species of parasitoids were reared from infested mines, with specimens belonging to the genus Chrysocharis Förster, 1856 (Hymenoptera: Eulophidae) and Itoplectis maculator (Fabricius, 1775) (Hymenoptera: Ichneumonidae). These species provided excellent control of leaf miner larvae, and out of 10 larvae, seven were found parasitized by the parasitoids and higher numbers of specimens were found with relative higher frequencies. [ABSTRACT FROM AUTHOR]

Most of the fire ants alien Solenopsis species, major pests of various horticulture crops. We have reported Solenopsis from crop fields and, through traditional taxonomy, these have been identified as Solenopsis geminata. The present investigation was performed to generate the DNA barcode of this species, for the elucidation of genetic structure collected from the Kalaburagi region. The generated DNA barcode of mitochondrial COI was deposited to the International Barcode of Life Database (BOLD: http://www.boldsystems.org). The barcode, along with its related information, are publicly available on the BOLD website with code MZ669889. It is the first such study from Kalaburagi region. Furthermore, DNA barcoding of fire ant's species accomplished from Maharashtra, Aurangabad (KC685060 & KC685063), Kerala, Malappuram (KP994543), Karnataka, Bangalore (KJ486555, KY000665 & JN886028) is in line with our findings. This study provides a detailed support for further wet lab study in support of traditional taxonomy. [ABSTRACT FROM AUTHOR]

Phylogenetic relationships of green-eyed skimmers, Macromia moorei was studied to estimate the evolutionary history among the members of family Corduliidae under order Odonata using the genetic methods of maximum probability, maximum parsimony, and Bayesian examination. Fifty-three DNA sequences and 3660 genetic characters/loci were used in the present study. Genetic distance of every nucleotide sequence was sorted from 0.69 to 98.10%. The outcomes of the combined COI + 16S DNA sequences revealed topology/phylogram through generally stronger bootstrap provision than evaluation of each gene in single analysis. Molecular analyses based on mitochondrial DNA yielded the position of M. moorei as a sister species of M.ampigena in family Corduliidae. [ABSTRACT FROM AUTHOR]

This study was conducted in the six municipalities of Apayao Province, namely, Luna, Pudtol, Flora, Conner, Kabugao, and Calanasan. This aimed to survey and characterize the edible fruit and ethnomedicinal trees in the forest landscape of Apayao province. It determined the geographical location, morphological characteristics, ecological status, DNA sequencing, phytochemical contents, uses, and threats of the edible and ethnomedicinal trees in the forests of Apayao. The methods used were qualitative and quantitative research. Fifteen (15) edible and 10 ethnomedicinal trees were surveyed with sixteen (16) families were identified. Out of 25 edible and ethnomedicinal trees, the conservation status is endangered, threatened, rare, vulnerable, and least concerned. Out of 25 edible and ethnomedicinal fruit trees, most are with identity results that range from 93 to 100% identity. Flavonoids, tannins, and sapotin compounds are mostly present in edible and ethnomedicinal trees. The community members are using 15 different ethnomedicinal trees to address 32 health-related conditions. The results of the phytochemical analyses provide support evidence to support the traditional uses of ethnomedicinal trees. All surveyed trees are susceptible to pests, diseases, and destruction brought by natural phenomena such as the effect of climate change. A policy recommendation for the conservation and protection of edible and ethnomedicinal trees is then proposed. [ABSTRACT FROM AUTHOR]

Studying the early stages of the introduction of non-indigenous species (NIS) is crucial as it allows immediate management actions aimed at preventing NIS spread at a time when these actions are more likely to be effective. Recent species introductions present unique opportunities to study key aspects of the invasion process. However, comprehensive information on how and when NIS are first introduced remains rare. We assessed the characteristics of the introduction of a member of the widespread Pyura stolonifera species group (Chordata, Tunicata) in Europe. These ascidians are important bioengineer species that can attain amongst the highest benthic biomass per surface area ever reported. We collected introduced individuals on the coast of the northwestern Iberian Peninsula, an area with high shipping traffic and one of the world's most important mussel farming regions. The specimens were analysed using taxonomic and genetic tools. Both field surveys and taxonomic analysis showed that one of the African members of the species group, Pyura herdmani, has recently been introduced to Europe and, although it does not yet form the large aggregates found in the native range, it is already well-established. Genetic data revealed that only the northwest African lineage of P. herdmani has been introduced to Europe. Considering the low dispersal capabilities of the early life-history stages of P. herdmani, the introduction of this species into Europe can only be explained via human mediated transport of NIS. In order to prevent any detrimental effects on both native biota and/or economically-important anthropogenic activities, regular monitoring of recent NIS introductions is urgently required. [ABSTRACT FROM AUTHOR]

Introduction: The genus Acronema, belonging to Apiaceae, includes approximately 25 species distributed in the high-altitude Sino-Himalayan region from E Nepal to SW China. This genus is a taxonomically complex genus with often indistinct species boundaries and problematic generic delimitation with Sinocarum and other close genera, largely due to the varied morphological characteristics. Methods: To explore the phylogenetic relationships and clarify the limits of the genus Acronema and its related genera, we reconstructed a reliable phylogenetic framework with high support and resolution based on two molecular datasets (plastome data and ITS sequences) and performed morphological analyses. Results: Both phylogenetic analyses robustly supported that Acronema was a non-monophyletic group that fell into two clades: Acronema Clade and East-Asia Clade. We also newly sequenced and assembled sixteen Acronema complete plastomes and performed comprehensively comparative analyses for this genus. The comparative results showed that the plastome structure, gene number, GC content, codon bias patterns were high similarity, but varied in borders of SC/IR and we identified six different types of SC/IR border. The SC/IR boundaries of Acronema chienii were significantly different from the other Acronema members which was consistent with the type VI pattern in the genus Tongoloa. We also identified twelve potential DNA barcode regions (ccsA, matK, ndhF, ndhG, psaI, psbI, rpl32, rps15, ycf1, ycf3, psaI-ycf4 and psbM-trnD) for species identification in Acronema. The molecular evolution of Acronema was relatively conservative that only one gene (petG) was found to be under positive selection (ω = 1.02489). Discussion: The gene petG is one of the genes involved in the transmission of photosynthetic electron chains during photosynthesis, which plays a crucial role in the process of photosynthesis in plants. This is also a manifestation of the adaptive evolution of plants in high-altitude areas to the environment. In conclusion, our study provides novel insights into the plastome adaptive evolution, phylogeny, and taxonomy of genus Acronema. [ABSTRACT FROM AUTHOR]

The genus Amomum includes over 111 species, 6 of which are widely utilized as medicinal plants and have already undergone taxonomic revision. Due to their morphological similarities, the presence of counterfeit and substandard products remains a challenge. Accurate plant identification is, therefore, essential to address these issues. This study utilized 11 newly sequenced samples and extensive NCBI data to perform molecular identification of the six medicinal "Doukou" species. The plastomes of these species exhibited a typical quadripartite structure with a conserved gene content. However, independent variation shifts of the SC/IR boundaries existed between and within species. The comprehensive set of genetic sequences, including ITS, ITS1, ITS2, complete plastomes, matK, rbcL, psbA-trnH, and ycf1, showed varying discrimination of the six "Doukou" species based on both distance and phylogenetic tree methods. Among these, the ITS, ITS1, and complete plastome sequences demonstrated the highest identification success rate (3/6), followed by ycf1 (2/6), and then ITS2, matK, and psbA-trnH (1/6). In contrast, rbcL failed to identify any species. This research established a basis for a reliable molecular identification method for medicinal "Doukou" plants to protect wild plant resources, promote the sustainable use of medicinal plants, and restrict the exploitation of these resources. [ABSTRACT FROM AUTHOR]

Aotearoa (New Zealand) is a biodiversity hotspot for temperate invertebrate taxa and home to high levels of endemicity. However, our knowledge of species‐level diversity and phylogeny of endemic New Zealand Onychophora (velvet worms) is at present limited. Here, we use mitochondrial cytochrome c oxidase subunit I (COI) barcoding to assess the extent of species diversity for the two velvet worm genera found in New Zealand, the ovoviviparous and endemic Peripatoides and the oviparous Ooperipatellus, found in Australia and New Zealand. Our results reveal that the estimated number of species of both genera in New Zealand is greater than currently described. We estimate there are between 13 and 67 species of Peripatoides and between 16 and 21 species of Ooperipatellus endemic to New Zealand. This is a stark increase from the two currently described New Zealand species of Ooperipatellus and previous work that has identified 10 species within Peripatoides. Our exploration of climatic variables shows that individuals of Ooperipatellus are predominantly found in wet, cool environments and Peripatoides are found across relatively drier, warmer habitats. We also generate ecological niche models to provide initial predictions of the distribution of climatically suitable habitats for each genus across New Zealand. [ABSTRACT FROM AUTHOR]

Snow Lotus Seed (SLS), esteemed for its nutritional and market value, faces challenges of authentication due to the absence of appropriate testing standards and methods. This results in frequent adulteration of SLS sourced from Gleditsia sinensis (G. sinensis) with other plant seeds endosperm. Traditional chloroplast DNA barcoding methods are inadequate for species identification due to the absence of chloroplasts in G. sinensis seeds endosperm. In this study, the homology of 11 ITS genes among 6 common Gleditsia species was analyzed. Universal primers suitable for these species were designed and screened. A DNA barcoding method for distinguishing SLS species was developed using Sanger sequencing technology, leveraging existing GenBank and Barcode of Life Data System (BOLD) databases. Optimized sample pretreatment facilitated effective DNA extraction from phytopolysaccharide-rich SLS. Through testing of commercial SLS products, the species origin has been successfully identified. Additionally, a novel instance of food fraud was uncovered, where the Caesalpinia spinosa endosperm was used to counterfeit SLS for the first time. The study established that the developed DNA barcoding method is effective for authenticating SLS species. It is of great significance for combating food fraud related to SLS, ensuring food safety, and promoting the healthy development of the SLS industry. [ABSTRACT FROM AUTHOR]

Acanthopsoides molobrion is a freshwater previously known to be spread across Malaysia and Borneo. This study aimed to record for the first time the occurrence of the A. molobrion in Bangka Island, Indonesia, and update the geographic distribution of this species. This new record of freshwater fish provides an important contribution to the comprehension of the biogeography of the species. The purpose sampling approach was employed for this study to collect specimens. The species were then identified morphologically by the application of morphometric and meristic methods, and molecularly through the employ of DNA Barcoding. On 20th January 2023, two A. molobrion specimens were collected from the Bumang Kemuja River, Bangka Island using a fish trap. The new record of A. molobrion found here is the southernmost record for this species, expanding its geographic distribution. In addition, the latest record site is about 500 km south of the nearest locality in Peninsular Malaysia, and about 750 km southwest of the nearest locality in Borneo. The new record of A. molobrion has expanded the species' recorded distribution range, which has added to our understanding of this species. Furthermore, we present an updated record for the A. molobrion DNA sequence based on the COI gene. This sequence is the first DNA Barcode to Indonesia. Subsequently, the DNA sequence was registered into NCBI Genbank with the access code OR144414. This DNA barcode will be used as a standard for identifying A. molobrion and will aid in DNA and biotechnology-based studies in the future. [ABSTRACT FROM AUTHOR]

Astragalus L. is one of the largest genera, including over 3000 taxa worldwide. In the present study, we found two new records of A. teskhemicus Sytin and Shaulo and A. testiculatus Pall. in western Mongolia. We provided a morphological revision and DNA barcoding (ITS) of two new records along with that of the most similar Astragalus species including A. ammodytes Pall., A.gubanovii N. Ulziykh., A. hypogaeus Ledeb., A. junatovii Sanchir. Furthermore, we made detailed photo illustrations and wild photos for new records and four species based on our field observations, respectively. The general distribution, taxonomic notes, type information, habitats, and species examined are given. [ABSTRACT FROM AUTHOR]

Rice cultivation is one of the most important agro-economic activities in many countries, and the correct seed storage between production cycles is essential for crop success. In Guinea-Bissau, mangrove swamp rice (MSR) is a highly productive rice cropping system, thus providing surplus for sale. Depending on storage conditions, rice grains may present moisture, insects, or the development of fungi that might affect crop productivity. Considering different rice varieties and storage conditions, samples were collected from 30 farmers within 13 villages across the main MSR-producing regions. Stored rice was used to isolate and identify seed-borne fungi through DNA barcoding, to test aflatoxin content, and to evaluate seed germination rates. Polyethylene bags (the container), raised wooden platforms, and storage rooms (the structures) are the most used facilities. Hermetic containers were recorded mainly in Oio. A total fungal richness of 18 genera was found, and 16 different species were identified. The most represented fungal genera are Aspergillus, Curvularia, and Bipolaris. Despite the presence of aflatoxin-producing fungal samples, they did not present concerning levels for human consumption, and the germination rate was not affected regardless of storage structures. These results provide a baseline on fungi occurrence in stored MSR seeds from traditional facilities in Guinea-Bissau. [ABSTRACT FROM AUTHOR]

Rhubarb is a significant medicinal herb in China. Its adulteration or fabrication is common in the market. Consequently, it is necessary to establish a comprehensive identification method to accurately identify genuine rhubarb and its adulterants. In this study, the sequences of chloroplast genes rps3-rpl22 and rpl16 from three genuine rhubarbs (Rheum tanguticum, Rh. palmatum and Rh. officinale) and their adulterants (Rumex japonicus and Rumex spp.) were amplified, sequenced and subjected to genetic analyses. The genetic distances for rps3-rpl22 and rpl16 between genuine rhubarbs and their adulterants showed that there was an evident barcoding gap, which allowed the adulterants to be distinguished from the genuine rhubarbs, as demonstrated by a neighbor joining tree. Additionally, Rh. officinale could be distinguished from the other two genuine rhubarbs. The anthraquinone, sennoside, polysaccharide and protein contents were analyzed in seven rhubarbs using high-performance liquid chromatography and ultraviolet spectrophotometry. Cluster and principal component analyses results showed that Rh. tanguticum and Rh. palmatum could be effectively distinguished. The study suggests that DNA barcoding based on rps3-rpl22 and rpl16 sequences coupled with multiple-indicator quantification can be successfully applied to identify rhubarb species and distinguish among the three genuine rhubarbs, and this can provide a scientific foundation for rhubarb quality assurance. [ABSTRACT FROM AUTHOR]

The utility of a universal DNA 'barcode' fragment (658 base pairs of the Cytochrome C Oxidase I [COI] gene) has been established as a useful tool for species identification, and widely criticized as one for understanding the evolutionary history of a group. Large amounts of COI sequence data have been produced that hold promise for rapid species identification, for example, for biosecurity. The fruit fly tribe Dacini holds about a thousand species, of which 80 are pests of economic concern. We generated a COI reference library for 265 species of Dacini containing 5601 sequences that span most of the COI gene using circular consensus sequencing. We compared distance metrics versus monophyly assessments for species identification and although we found a 'soft' barcode gap around 2% pairwise distance, the exceptions to this rule dictate that a monophyly assessment is the only reliable method for species identification. We found that all fragments regularly used for Dacini fruit fly identification >450 base pairs long provide similar resolution. 11.3% of the species in our dataset were non‐monophyletic in a COI tree, which is mostly due to species complexes. We conclude with recommendations for the future generation and use of COI libraries. We revise the generic assignment of Dacus transversus stat. rev. Hardy 1982, and Dacus perpusillus stat. rev. Drew 1971 and we establish Dacus maculipterus White 1998 syn. nov. as a junior synonym of Dacus satanas Liang et al. 1993. [ABSTRACT FROM AUTHOR]

The Ib River is an important tributary of the Mahanadi River. The aim of the present study was to establish a reliable barcoding reference database for fish in Ib River. A total of 134 specimens, belonging to 42 species within 32 genera, 18 families, and 11 orders, were subjected to PCR amplification of the target region of mitochondrial genome and sequenced. The average length of the 134 barcodes was 648 bp. The average K2P distances within species, genera, and families were 0.13, 9.94, and 16.79%, respectively. The average interspecific distance was approximately 76.46 times higher than the mean intraspecific distance. The Barcode Index Number (BIN) discordance report showed that 134 specimens represented 42 BINs, of which 13 were singletons, 29 were taxonomically concordant while no species taxonomically discordant in our dataset. The barcode gap analysis demonstrated that 100% of the fish species examined could be discriminated by DNA barcoding. These results provide new insights into fish diversity in the Ib River and can provide a basis for the development of further studies on fish fauna. [ABSTRACT FROM AUTHOR]

Valettietta Lincoln & Thurston, 1983 (Amphipoda: Alicelloidea) is an infrequently sampled genus of scavenging amphipod, with a known bathymetric range from 17–5467 m encompassing a variety of habitats from anchialine caves to abyssal plains. Molecular systematics studies have uncovered cryptic speciation in specimens collected from the abyssal Pacific, highlighting uncertainty in the description of Valettietta anacantha (Birstein & Vinogradov, 1963). Here, we apply an integrative taxonomic approach and describe two new species, Valettietta trottarum sp. nov. and Valettietta synchlys sp. nov. collected at abyssal depths in the Clarion-Clipperton Zone, Pacific Ocean. Both species can be distinguished by characters of the gnathopods, uropod 3, and the inner plate of the maxilliped. Further, molecular phylogenetic analyses of two mitochondrial (16S rDNA and COI) and two nuclear (Histone 3 and 28S rRNA) regions found both new species to form well-supported clades and allowed us to re-identify previously published records based on genetic species delimitation. The biogeography of Valettietta is discussed in light of these re-evaluated records, and a new taxonomic key to the genus is provided. These new taxa highlight the strength of applying an integrated taxonomic approach to uncover biodiversity, which is critical in regions being explored for potential industrial purposes. [ABSTRACT FROM AUTHOR]

Checkpoint inhibition (CPI) therapy and adoptive cell therapy with autologous tumor-infiltrating lymphocytes (TIL-based ACT) are the two most effective immunotherapies for the treatment of metastatic melanoma. While CPI has been the dominating therapy in the past decade, TIL-based ACT is beneficial for individuals even after progression on previous immunotherapies. Given that notable differences in response have been made when used as a subsequent treatment, we investigated how the qualities of TILs changed when the ex vivo microenvironment of intact tumor fragments were modulated with checkpoint inhibitors targeting programmed death receptor 1 (PD-1) and cytotoxic Tlymphocyte-associated protein 4 (CTLA-4). Initially, we show that unmodified TILs from CPI-resistant individuals can be produced, are overwhelmingly terminally differentiated, and are capable of responding to tumor. We then investigate these properties in ex vivo checkpoint modulated TILs finding that that they retain these qualities. Lastly, we confirmed the specificity of the TILs to the highest responding tumor antigens, and identified this reactivity resides largely in CD39+CD69+ terminally differentiated populations. Overall, we found that anti-PD-1 will alter the proliferative capacity while anti-CTLA4 will influence breadth of antigen specificity. [ABSTRACT FROM AUTHOR]

Ladybirds (Coleoptera: Coccinellidae) are important predatory insects found in many croplands, but their patterns of diversity and assemblage in diverse crop management practices remain understudied, especially in southeastern Asia. Their existence denotes a crucial need to update the diversity and assemblage pattern in diverse crop management practices. This study aims to (i) delimit ladybird species through DNA barcodes and (ii) compare the abundance of different ladybird taxa from different crop management practices. A total of 2260 ladybirds were collected and barcoded resulting in 12 species representing four subfamilies (Coccidulinae, Coccinellinae, Epilachninae, and Scymninae). Three predatory species dominated and were top contributors to the dissimilarity average for different crop management practices, i.e., Coccinella transversalis, Micraspis discolor, and Cheilomenes sexmaculata. Even though the effect of different crop management practices on ladybird abundance was insignificant (Kruskal Wallis, p-value > 0.05), their diversity significantly varied across different practices (diversity t-test, p-value < 0.05). Organic monocrop resulted in a higher value of Shannon index (H'), and richness than other management practices. Furthermore, monocrops comprised higher ladybird diversity than multicrops. While the species assemblage was not distinct (ANOSIM, p-value > 0.05), a variation in assemblage composition and spatial distribution concerning the different crop management employed (NMDS, stress value = 0.12) was observed. The hierarchical dendrogram distinguished six clusters of ladybirds between organic and conventional management practices. More explorations are required to uncover the various effects of crop management practices on ladybirds' fitness and survival in different landscapes. [ABSTRACT FROM AUTHOR]

An unidentified ant species was discovered in the heart of New York City in 2011, which garnered national headlines and created the memorable nickname "ManhattAnt." New York City is one of the oldest and largest cities in North America and has been the site of introduction for some of North America's most damaging invasive pests. Nevertheless, there has been little follow up research on the ManhattAnt since its discovery, and it has yet to be formally identified. Here we use genetic and morphological approaches to confirm the identity of the ManhattAnt and characterize its introduced range and colony structure. Results from genetic analyses placed the ManhattAnt within the Lasius emarginatus species complex, and morphological comparisons ruled out closely related species to identify the ManhattAnt as the European ant L. emarginatus with 93–99% certainty depending on the nest sample. Since its initial discovery in the region, the ManhattAnt has become one of the most common ants in New York City and has spread at a rate of 2 km/yr into New Jersey and onto Long Island. Based on the climate it inhabits within its native range, L. emarginatus could expand to cover much of the eastern United states from Massachusetts to Georgia. Although many successful invasive ants display supercoloniality with little aggression between neighboring nests, we found no evidence that L. emarginatus colonies in New York City are supercolonial. Continued monitoring of L. emarginatus is warranted, as it has been increasingly reported as an indoor pest and is known to form mutualisms with honeydew producing pests of street trees. [ABSTRACT FROM AUTHOR]

The release of ornamental pets and associated pathogens outside their native range might directly or indirectly impact the recipient community. In temperate regions, e.g., central Europe, feral freshwater species of tropical and sub-tropical origins are mainly constrained to thermally polluted waters and thermal springs. However, species with high environmental plasticity and reproduction rates, such as the shrimp Neocaridina davidi, may adapt to colder water regimes over time. A widening thermal niche may eventually overcome thermal barriers, further expanding the range and enhancing transmission opportunities for host generalist parasites. This study assesses the observed (field observations) and theoretical (species distribution models) range expansion of N. davidi and associated parasites in Europe. We report three newly established N. davidi populations from thermally polluted waters in central Europe (Germany, Hungary, and Slovakia) and provide further evidence of its range expansion into colder environments. Species distribution models predict thermally suitable habitats in the Mediterranean and a foreseeable expansion into Western Europe and the Balkans by 2050. We confirm the presence of the microsporidian parasite Ecytonucleospora hepatopenaei in feral N. davidi populations across Europe and expand the list of microsporidians found in this host from two to four. Furthermore, we provide the first evidence of parasite spillover from/to the invasive crayfish Procambarus clarkii, suggesting that parasite exchange with native biota might be possible. Such possibility, coupled with an ongoing range expansion of N. davidi bolstered by human-mediated introductions and climate change, will likely exacerbate the impact on native biota. [ABSTRACT FROM AUTHOR]

A new, distinctively short‐bodied giraffe catfish of Parauchenoglanis is described from the Ndzaa River, a small left‐bank tributary of the Mfimi‐Lukenie basin in the Central basin of the Congo River in the Democratic Republic of the Congo. The new species can be distinguished from all congeners by having 29 or fewer (vs. 33 or more) total vertebrae. It can further be distinguished from all congeners, except Parauchenoglanis zebratus Sithole et al., 2023 and Parauchenoglanis ngamensis (Boulenger 1911), by having 13 or 14 (vs. 16 or more) pre‐anal vertebrae. The species is endemic to the Mfimi River basin, where it has been collected mainly in blackwater tributaries. [ABSTRACT FROM AUTHOR]

Backgroud: The Northeast India, being part of two global biodiversity hotspot namely the Indo-Burma and Eastern Himalayan Hotspots supports a wide variety of rich aquatic biodiversity including fishes. The family Danionidae is a widely diverse group inhabiting the upper colder stretches of river although few are abundant in the lower stretches. The persisting similarity in the morphological appearance and body colouration within the members of this family seeks an integrated method to identify the species correctly. Methods and results: In the present study, the mt-DNA barcode was generated for correct identification and confirmation of the species. A total of nine mitochondrial cytochrome c oxidase subunit I gene sequences were generated for each species under the study. The pairwise distance values ranged from 0.09 to 9.11% within species and 9.06–32.71% between species. A neighbour-joining tree was constructed based on the Kimura 2 parameter model. Two major groups were observed where Danioninae formed a sister group to the Chedrinae and Rasborinae. Conclusion: The present study is a preliminary work to document and identify the species under the family Danionidae from Brahmaputra basin, Assam, using molecular tools and establish the phylogenetic relationship. [ABSTRACT FROM AUTHOR]

Background: The Anopheles hyrcanus group is distributed throughout the Oriental and Palaearctic regions and can transmit diseases such as malaria, Japanese encephalitis virus, and filariasis. This investigation marks the inaugural comprehensive study to undertake a phylogenetic analysis of the constituents of this malaria vector group in the northeastern region of Iran, juxtaposed with documented occurrences from different areas within Iran and worldwide. Methods: Mosquitoes were collected using various methods from nine different locations in Golestan province from April to December 2023. The collected mosquitoes were identified morphologically using valid taxonomic keys. DNA was isolated using the Sambio™ Kit. COI and ITS2 primers were designed using Oligo7 and GeneRunner. PCR and purification were performed with the Qiagen kit. Subsequently, sequencing was carried out at the Mehr Mam GENE Center using an Applied Biosystems 3730XL sequencer. The nucleotide sequences were then analyzed and aligned with GenBank data using BioEdit. Kimura 2-parameter was Utilized for base substitutions. DNA models were selected based on AIC and BIC criteria. Bayesian and Maximum Likelihood trees were constructed, along with a haplotype network. Molecular diversity statistics computed using DnaSP software. Results: In this study, a total of 819 adult mosquitoes were collected. An. hyrcanus was the second most abundant species, predominantly found in Kalaleh and Turkman counties. The sequenced and edited COI and ITS2 sequences were deposited in GenBank under specific accession numbers. Phylogenetic analyses using ML, BI, and NJ methods confirmed a monophyletic lineage for An. hyrcanus with strong support. Molecular analysis of Iranian An. hyrcanus found 11 diverse haplotypes, with the COI gene displaying low diversity. The ITS2 gene revealed two clades - one associating with Iran, Europe, and Asia; the other originating from southwestern Iran. The haplotype network showed two main groups - one from southwest Iran and the other from north Iran. Iran exhibited six distinct haplotypes, while Turkey showcased the highest diversity. Conclusions: An. hyrcanus in southwestern Iran exhibits a distinct haplogroup, suggesting possible subspecies differentiation. Additional studies are required to validate this phenomenon. [ABSTRACT FROM AUTHOR]

Specimen identification at the species level is a critical challenge for understanding community structure and conserving biodiversity. The use of mitochondrial DNA barcodes, in addition to morphology, has proven to be a useful tool for earthworm identification, but it has also raised difficulties. Thus, approaches to delineating molecular operational taxonomic units (MOTUs) from cytochrome c oxidase subunit 1 (COI) or 16S rDNA sequence data often reveal more mitochondrial lineages than morphospecies, raising the question of whether these MOTUs should be used as taxonomic entities in community structure studies. Here, we used a newly acquired dataset of 576 COI barcodes of earthworms from Metropolitan France that were clustered in 36 MOTUS, corresponding to 21 morphospecies. We also incorporated data gathered from the literature to investigate this question. In order to match our MOTUs with already-described mitochondrial lineages, we downloaded reference sequences from the GenBank and BOLD platforms. In light of the difficulties encountered in recovering these sequences, we recommend that any new mitochondrial lineage described in a study be named consistently with previous works. Next, we analyzed the biological, ecological, and molecular data available in the literature on the different mitochondrial lineages that matched our MOTUs in order to determine if there was a consensus for species delimitation. Although the study specimens mainly belong to the Lumbricidae, which is one of the most studied families of earthworms, the data are often missing to determine if the MOTUs correspond to different species. Pending revision of the taxonomy, MOTUs for which mitochondrial divergence has been confirmed by morphological differences should be considered distinct taxonomic entities in community structure studies. In the absence of morphological differences and pending more data, we propose to distinguish for these analyses, within sexually reproducing morphospecies, the MOTUs for which mitochondrial divergence has been confirmed by multilocus nuclear data, while in the case of reproduction by parthenogenesis, ecological differences between MOTUs are necessary to consider them as different taxonomic entities. [ABSTRACT FROM AUTHOR]

[Objective] To identify the female fly intercepted from imported mung bean by morphological classification and DNA barcoding techniques. [Method] A female fly which were intercepted from mung bean on an container from Uzbekistan at Henan Port on February 23, 2023. And it was identified by using morphological classification and DNA barcoding techniques, and it was compared with its similar species Pollenia rudis. [Result] The intercepted fly was identified as Pollenia pediculate, female. It was intercepted for the first time at Henan Port. [Conclusion] Flies could be identified rapidly and accurately by using DNA barcoding and morphological classification. [ABSTRACT FROM AUTHOR]

This study investigated the diversity, composition and distribution patterns of polychaete macrofauna inhabiting unconsolidated sediments on the continental shelf of southern Namibia. During the austral summer of 2021, 910 Van Veen grab samples were collected from 91 sites in water depths ranging between 43 and 146 m. All benthic macrofauna (> 1 mm) were extracted, identified, enumerated and weighed. Polychaetes were the most abundant taxon, equating to 66% of total abundance and 37% of total biomass. A total of 83 712 polychaete specimens comprising 112 species, 80 genera and 33 families were collected. Several taxa identified were listed as putative cosmopolitans (19 taxa) or have known wide local distributions (10 taxa). Voucher specimens were collected for DNA barcoding to improve reference sequence libraries for the region. Hierarchical cluster analyses using both abundance and biomass data were undertaken to determine spatial distribution patterns in polychaete communities. Both datasets yielded similar results with polychaete assemblages divided into inshore and offshore communities, that were further subdivided at a local scale. Investigation of physical and chemical drivers suggested that polychaete communities in southern Namibia are structured to varying degrees by water depth, latitude, sediment composition, redox potential and organic content. Deeper stations comprised the highest diversity of polychaeta fauna. Species adapted to hypoxic conditions (e.g. Sigambra parva, Pararionospio pinnata, Diopatra cf. monroi and Nepthys cf. hombergii) dominated an area known as the mudbelt, where organically enriched silts and clays originating from the Orange River are deposited on the mid-shelf between the 40 and 120 m isobaths. [ABSTRACT FROM AUTHOR]

This article explores the use of DNA barcoding as a tool for studying the biodiversity and genetic relationships of local cultivars of Zea and Trifolium plants in Egypt. The study collected samples from field crops and analyzed the DNA sequences using different markers. The results provide insights into the genetic diversity and relationships among different cultivars. The article also includes various scientific articles and publications related to DNA barcoding and genetic diversity in plant species, focusing on specific plant species such as clover, teosinte, and jute. The study's results, obtained from the GenBank database, offer valuable information for researchers studying these plant species and their genetic relationships. [Extracted from the article]

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]

Several individuals of an unidentified Goodyera taxon were discovered in Tomakomai City, Hokkaido Prefecture, Japan. Detailed morphological analysis suggests that this taxon is a natural hybrid between Goodyera schlechtendaliana and Goodyera henryi, based on characteristics such as leaf margins, texture, venation patterns, rachis length, hair morphology on the ovary, and the shapes of the lip apex and rostellum. Supporting this, molecular data from genome-wide markers using next-generation sequencing (MIG-seq data) indicate that G. xkazumae is a hybrid between G. schlechtendaliana and G. henryi. Neighbor-Net phylogenetic analysis shows G. xkazumae occupies an intermediate position between the putative parent species. STRUCTURE analysis further confirms that G. xkazumae comprises genetic components from both species. Therefore, we conclude that this taxon is a natural hybrid based on comprehensive morphological and molecular evidence. [ABSTRACT FROM AUTHOR]

The hemiparasitic tribe Cymbarieae (Orobanchaceae) plays a crucial role in elucidating the initial stage of the transition from autotrophism to heterotrophism. However, the complete chloroplast genome of the type genus Cymbaria has yet to be reported. In addition, the traditional Mongolian medicine Cymbaria daurica is frequently subjected to adulteration or substitution because of the minor morphological differences with Cymbaria mongolica. In this study, the complete chloroplast genomes of the two Cymbaria species were assembled and annotated, and those of other published 52 Orobanchaceae species were retrieved for comparative analyses. We found that the Cymbaria chloroplast genomes are characterized by pseudogenization or loss of stress‐relevant genes (ndh) and a unique rbcL–matK inversion. Unlike the high variability observed in holoparasites, Cymbaria and other hemiparasites exhibit high similarity to autotrophs in genome size, guanine–cytosine (GC) content, and intact genes. Notably, four pairs of specific DNA barcodes were developed and validated to distinguish the medicinal herb from its adulterants. Phylogenetic analyses revealed that the genus Cymbaria and the Schwalbea–Siphonostegia clade are grouped into the tribe Cymbarieae, which forms a sister clade to the remaining Orobanchaceae parasitic lineages. Moreover, the diversification of monophyletic Cymbaria occurred during the late Miocene (6.72 Mya) in the Mongol–Chinese steppe region. Our findings provide valuable genetic resources for studying the phylogeny of Orobanchaceae and plant parasitism, and genetic tools to validate the authenticity of the traditional Mongolian medicine "Xinba." [ABSTRACT FROM AUTHOR]

Torinido‐shoujoubae, as it is called in Japanese, is a flightless Drosophila sp. that is sold commercially in Japan. This Drosophila sp. is often used as feeds for model organisms such as reptiles and spiders. There is no scientific name provided for the fruit fly that is known as Torinido‐shoujoubae, as well as any historical background or data behind this species. There has been a previous study that was conducted through morphological characteristics analysis of the body as well as the male copulatory organ and has been estimated as Drosophila hydei. The objective of this study was to determine the species of this unidentified fly known as Torinido‐shoujoubae based on a molecular evidence with a DNA barcoding. Samples were purchased from four separate suppliers to examine whether there are any differences between them. COI regions were amplified using PCR and the sequenced results were aligned against two databases, NCBI and BOLD. Torinido‐shoujoubae samples provided from all suppliers were confirmed to be D. hydei. [ABSTRACT FROM AUTHOR]

Aim: A detailed, comparative DNA‐barcoding and morphospecies‐based overview of the vertical and horizontal distribution of Lake Ohrid's endemic Gammarus species flock is provided. Re‐evaluation of the time frame for the onset of the species flock and identification of events that putatively influenced diversification processes. Location: Lake Ohrid, Balkan Peninsula, Europe. Taxon: Gammarus species flock (Amphipoda, Crustacea). Methods: Extensive sampling and DNA barcoding of 600 individuals was carried out. DNA sequences were analysed using species delimitation methods, haplotype network reconstructions, Bayesian molecular dating and demographic analyses. The COI‐based species delimitation results were validated with nuclear 28S RNA data. Results: The species flock distribution has a weak horizontal but clear vertical structure. The diversity along bathymetric gradients correlates with temperature and salinity; and the highest diversity is observed in sublittoral and springs on the lake's shore. Two new molecular units representing putatively new species are revealed and supported also by the nuclear marker. The onset of flock radiation overlaps with the time of lake formation. The COI gene shows signs of positive selection and an acceleration in substitution rate. The demographic changes within the flock happened during the last thousands years. Main Conclusions: Distribution of the Gammarus species flock is vertically structured, reflecting habitat zonation. Parapatric speciation is one of the possible mechanisms behind the flock's diversification. Detection of new MOTUs suggests that the flock's diversity is still not fully revealed. Simultaneously, failure to collect three previously known MOTUs suggests a loss of gammarid diversity in the lake. This represents, together with the current threats to the lake ecosystem (i.e. climate changes, development of tourism), a clear call for conservation efforts. The speciation events and demographic changes within the flock relate presumably to glacial and postglacial water level changes and to the colonisation of new depth ranges and the associated springs. [ABSTRACT FROM AUTHOR]

Ultrasensitive protein identification is of paramount importance in basic research and clinical diagnostics but remains extremely challenging. A key bottleneck in preventing single-molecule protein sequencing is that, unlike the revolutionary nucleic acid sequencing methods that rely on the polymerase chain reaction (PCR) to amplify DNA and RNA molecules, protein molecules cannot be directly amplified. Decoding the proteins via amplification of certain fingerprints rather than the intact protein sequence thus represents an appealing alternative choice to address this formidable challenge. Herein, we report a proof-of-concept method that relies on residue-resolved DNA barcoding and composition code counting for amplifiable protein fingerprinting (AmproCode). In AmproCode, selective types of residues on peptides or proteins are chemically labeled with a DNA barcode, which can be amplified and quantified via quantitative PCR. The operation generates a relative ratio as the residue-resolved 'composition code' for each target protein that can be utilized as the fingerprint to determine its identity from the proteome database. We developed a database searching algorithm and applied it to assess the coverage of the whole proteome and secretome via computational simulations, proving the theoretical feasibility of AmproCode. We then designed the residue-specific DNA barcoding and amplification workflow, and identified different synthetic model peptides found in the secretome at as low as the fmol/L level for demonstration. These results build the foundation for an unprecedented amplifiable protein fingerprinting method. We believe that, in the future, AmproCode could ultimately realize single-molecule amplifiable identification of trace complex samples without further purification, and it may open a new avenue in the development of next-generation protein sequencing techniques. [ABSTRACT FROM AUTHOR]

Meretrix, a popular marine bivalve, features several species with significant ecological and commercial value, distributed in various marine habitats worldwide. Despite their importance, accurate global population estimates of these species are hindered by inadequate classification frameworks that often rely on external shell characteristics, which are susceptible to environmental influences. This limitation underscores the necessity for comprehensive research into the physiological and molecular attributes of these species for accurate identification. This study focused on identifying the local Meretrix species from Tarakan Island by analyzing their COI nucleotide sequences and comparing them with reference sequences in the BOLD and GenBank databases. The results demonstrate that the 14 specimens obtained with diverse morphological variations were found only within two major groups: Meretrix meretrix and Meretrix lusoria. These findings validate the efficacy of DNA barcoding as a tool for classifying marine bivalves, offering a robust framework for identifying unknown species, particularly in the Asian marine ecosystems. The study's approach and findings have broader implications for enhancing global marine biodiversity research and conservation strategies. [ABSTRACT FROM AUTHOR]

Mitragyna speciosa, commonly known as kratom, is a narcotic plant that is used for its unique mood‐enhancing and pain‐relieving effects. It is marketed throughout the United States as a 'legal high' and has gained popularity as an alternative to opioids. However, kratom's increasing involvement in accidental overdoses, especially among polydrug users, has prompted warnings from the Drug Enforcement Agency (DEA) and the Food and Drug Administration (FDA). Despite these warnings, kratom remains legal federally, although it is banned in six states. This legal disparity complicates monitoring and enforcement efforts in states where kratom is illegal. Common forensic techniques using morphology or chemical analysis are beneficial in some instances but are not useful in source attribution because most seized kratom is powdered and the alkaloid content of samples can vary within products, making sourcing unreliable. This study focused on developing a DNA barcoding method to access sequence variation in commercial kratom products. It evaluated the utility of one nuclear barcode region (ITS) and three chloroplast barcode regions (matK, rbcL, and trnH‐psbA) in assessing sequence variation across commercially available kratom products. Novel polymorphisms were discovered, and the ITS region showed the greatest variation between samples. Among the 15 kratom products tested, only two haplotypes were identified across the four barcoding regions. The findings highlight the potential of DNA barcoding as a forensic tool in the traceability and enforcement against illegal kratom distribution. Nonetheless, the limited haplotypic diversity points to a need for further development and expansion of the M. speciosa DNA sequence database. [ABSTRACT FROM AUTHOR]