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Multiple Salmonella enterica serovars and strains have been reported to be able to persist inside the foliar tissue of lettuce (Lactuca sativa L.), potentially resisting washing steps and reaching the consumer. Intraspecies variation of the bacterial pathogen and of the plant host can both significantly affect the outcome of foliar colonization. However, current understanding of the mechanisms underlying this phenomenon is still very limited. In this study, we evaluated the foliar fitness of 14 genetically barcoded S. enterica isolates from 10 different serovars, collected from plant and animal sources. The S. enterica isolates were vacuum-infiltrated individually or in pools into the leaves of three- to four-week-old lettuce plants. To estimate the survival capacity of individual isolates, we enumerated the bacterial populations at 0- and 10- days post-inoculation (DPI) and calculated their net growth. The competition of isolates in the lettuce apoplast was assessed through the determination of the relative abundance change of barcode counts of each isolate within pools during the 10 DPI experimental period. Isolates exhibiting varying apoplast fitness phenotypes were used to evaluate their capacity to grow in metabolites extracted from the lettuce apoplast and to elicit the reactive oxygen species burst immune response. Our study revealed that strains of S. enterica can substantially differ in their ability to survive and compete in a co-inhabited lettuce leaf apoplast. The differential foliar fitness observed among these S. enterica isolates might be explained, in part, by their ability to utilize nutrients available in the apoplast and to evade plant immune responses in this niche.

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]

Tardigrade diversity and distribution are enigmatic in most parts of the globe, and only some European countries can boast of a relatively well-studied water bear fauna. However, even these suffer from the lack of genetic data, which would substantiate faunistic data and make biogeographic comparisons easier. Denmark has never been intensively and systematically researched in this regard, thus a citizen science sampling of cryptogams (mosses, liverworts, and lichens) was launched in spring 2023, aiming at a comprehensive biodiversity survey across this insular country. Nearly 700 samples were selected out of 8.000 sent to NHMD, based on the quality of samples, representativeness of various regions of Denmark, and the type of substrate to allow unravelling of potential ecological associations between tardigrades and cryptogams. Importantly, a large fraction of morphological identifications was backed up by DNA barcode data based on ITS-2 (1001 sequences), and in some cases also on COI (93 sequences) and ITS-1 (22 sequences) molecular markers, which are recognised DNA fragments used in species delimitation. We quadruple the number of known Danish limno-terrestrial tardigrade species (55 spp. reported in this paper vs. 14 spp. reported in literature so far, most of which were contentious due to the insufficient knowledge on tardigrade taxonomy), demonstrating the power of integrative taxonomy. No fewer than nine spp. are new to science. This is the first case where tardigrade fauna of an entire country is examined both from morphological and DNA barcoding data perspective. [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]

Consumers are thought to select food resources based on their nutritional content. While laboratory experiments have explored this, the nutritional dynamics of invertebrate predators have been scarcely studied in the field given various methodological constraints. The intersection of these nutritional dynamics with predator traits is also poorly characterised, leading to many gaps in our understanding of how different predators forage and feed in natural systems. Here, we integrate dietary metabarcoding with prey macronutrient (protein, lipid and carbohydrate) content and abundance to assess how nutrients and predator traits (sex, life stage and taxonomy) interactively drive prey preferences in the field, using spider–prey interactions as a model system. Different spider genera, sexes and life stages had nutritionally distinct diets. Our analyses demonstrated disproportionate foraging (selection and avoidance) for prey rich in different macronutrients, with the nature of these relationships differing between spider taxa, life stages and sexes. This may be explained by niche differentiation among spider groups, driven by biases toward prey rich in different nutrients, or nutrient‐specific foraging in which individual spiders vary their nutritional preferences to redress deficits, although further evidence is required to confirm this. This insight into the nutritional dynamics of generalist invertebrate predators extends our understanding beyond lab‐based behavioural assays and provides a novel framework for other complex real‐world systems. [ABSTRACT FROM AUTHOR]

ABSTRACT Chironomidae, so‐called non‐biting midges, are considered key bioindicators of aquatic ecosystem variability. Data derived from morphologically identifying their chitinous remains in sediments document chironomid larvae assemblages, which are studied to reconstruct ecosystem changes over time. Recent developments in sedimentary DNA (sedDNA) research have demonstrated that molecular techniques are suitable for determining past and present occurrences of organisms. Nevertheless, sedDNA records documenting alterations in chironomid assemblages remain largely unexplored. To close this gap, we examined the applicability of sedDNA metabarcoding to identify Chironomidae assemblages in lake sediments by sampling and processing three 21–35 cm long sediment cores from Lake Sempach in Switzerland. With a focus on developing analytical approaches, we compared an invertebrate‐universal (FWH) and a newly designed Chironomidae‐specific metabarcoding primer set (CH) to assess their performance in detecting Chironomidae DNA. We isolated and identified chitinous larval remains and compared the morphotype assemblages with the data derived from sedDNA metabarcoding. Results showed a good overall agreement of the morphotype assemblage‐specific clustering among the chitinous remains and the metabarcoding datasets. Both methods indicated higher chironomid assemblage similarity between the two littoral cores in contrast to the deep lake core. Moreover, we observed a pronounced primer bias effect resulting in more Chironomidae detections with the CH primer combination compared to the FWH combination. Overall, we conclude that sedDNA metabarcoding can supplement traditional remain identifications and potentially provide independent reconstructions of past chironomid assemblage changes. Furthermore, it has the potential of more efficient workflows, better sample standardisation and species‐level resolution datasets. [ABSTRACT FROM AUTHOR]

Knowledge of global patterns of genetic diversity is essential for biodiversity conservation as this parameter describes the ability of a species to respond to environmental changes. Ichneumonoids parasitoid wasps are among the few taxa showing an anomalous latitudinal diversity gradient. Using the largest georeferenced molecular dataset for this group, we used a macrogenetics approach to examine latitudinal patterns and predictors of intraspecific genetic diversity. We calculated the mean nucleotide diversity of mitochondrial DNA barcode sequences at three geographic levels: grid cells, latitudinal bands and climatic zones. Nucleotide diversity values were consistently higher at northern temperate latitudes, peaking at 50°. We found a positive but weak relationship between intraspecific diversity and the latitude, between intra‐ and interspecific diversity, and a positive effect of the temperature range. Examining the spatial relationship between different levels of biodiversity and its drivers is particularly relevant considering climate change and its impact on species distribution. Yet, in insects, it has been challenging to integrate ecological, evolutionary and geographical components when analysing the processes leading to species richness gradients. [ABSTRACT FROM AUTHOR]

ABSTRACT It is a global priority to better manage the biosphere, but action must be informed by comprehensive data on the abundance and distribution of species. The acquisition of such information is currently constrained by high costs. DNA barcoding can speed the registration of unknown animal species, the most diverse kingdom of eukaryotes, as the BIN system automates their recognition. However, inexpensive sequencing protocols are critical as the census of all animal species is likely to require the analysis of a billion or more specimens. Barcoding involves DNA extraction followed by PCR and sequencing with the last step dominating costs until 2017. By enabling the sequencing of highly multiplexed samples, the Sequel platforms from Pacific BioSciences slashed costs by 90%, but these instruments are only deployed in core facilities because of their expense. Sequencers from Oxford Nanopore Technologies provide an escape from high capital and service costs, but their low sequence fidelity has, until recently, constrained adoption. However, the improved performance of its latest flow cells (R10.4.1) erases this barrier. This study demonstrates that a MinION flow cell can characterise an amplicon pool derived from 100,000 specimens while a Flongle flow cell can process one derived from several thousand. At $0.01 per specimen, DNA sequencing is now the least expensive step in the barcode workflow. [ABSTRACT FROM AUTHOR]

Although new data and methodologies are gaining prominence in the field of taxonomy, the vast majority of terrestrial isopod species are still described solely based on morphology. The genus Benthana was recently revised and currently comprises 30 species found in South America. However, the broad geographic distribution of some species, coupled with the presence of few diagnostic characters, raises doubts about the validity of the described species. This study aimed to test the applicability of DNA Barcoding to assess the congruence between morphological and molecular identification of Benthana species. We analyzed 72 individuals belonging to 13 species from the scientific collection of the UFRGS Crustacean Laboratory. We obtained 24 sequences from ten Benthana species. The molecular phylogenies demonstrated that Benthana forms a paraphyletic group. Even at the species level, paraphyletic taxa were observed. Species delimitation methods revealed the presence of eight molecular entities within Benthana. Regarding genetic distances, we observed low intra-specific distances and significantly high inter-specific distances. Our results show that molecular analysis does not support the current taxonomy of Benthana. Therefore, a reassessment of the genus systematics based on integrative taxonomy is warranted. [ABSTRACT FROM AUTHOR]

Astroblepus species, commonly known as Andean climbing catfish, exhibit a unique challenge in species delimitation, leading to ongoing taxonomic debates. Here we report data on Astroblepus mindoensis, a vulnerable species endemic to Ecuador, obtained by an integrative approach that includes cytogenetic analysis, molecular identification of the specimens, and recording of morphological and morphometric characters useful for species diagnosis. Thus, this study aimed to associate the karyotype data of the specimens analyzed with morphological and molecular characters, improving and expanding the existing taxonomic information, thus contributing to the systematics of the species. Our morphology results, unlike Regan's original description, which is brief and ambiguous, provide a more detailed morphometric and meristic description. Molecular phylogenetic reconstruction and genetic distance based on a fragment of the cytochrome c oxidase subunit I (COI) showed that our samples constitute a well‐supported and monophyletic clade within the A. grixalvii species complex. The cytogenetic analysis identified distinct chromosomal markers, including a single cluster of major ribosomal genes (on chromosome pair 3) and of minor ribosomal genes (on chromosome pair 12) with their localization differing from those reported in other Astroblepus species analyzed. Additionally, the presence of a heteromorphic chromosome pair in males suggests the presence of an XX/XY sex‐determination system that has not been identified in other congeneric species. Further investigation is necessary to determine if these chromosomes are associated with the accumulation of repeated sequences, as typically occurs with sex chromosomes, and to assess their presence in other species of the genus. [ABSTRACT FROM AUTHOR]

Zooplankton monitoring is important for understanding their population dynamics and life history, ecosystem health, and environmental changes. Compared with traditional morphological identification, environmental DNA (eDNA) analysis allows for more sensitive and efficient monitoring of zooplankton diversity. Previous eDNA studies have primarily used metabarcoding approaches to reveal their richness and composition, whereas its performance in predicting zooplankton abundance remains understudied. We conducted water and bulk sampling in Lake Biwa, Japan, showing that the number of sequence reads by metabarcoding moderately correlated with eDNA concentrations estimated by quantitative real-time PCR (qPCR). In addition, the eDNA read number was significantly related to cladoceran and copepod abundance estimated by microscopy sorting, although there remained too much uncertainty in the read-abundance relationship. Moreover, there was a significant difference in species composition between eDNA metabarcoding and sorting. Although our results indicated the potential applicability of eDNA metabarcoding for quantifying multiple zooplankton abundance, several methodological validations in eDNA metabarcoding would also be required to optimize its performance in the future. [ABSTRACT FROM AUTHOR]

Phytoplankton have short generation times, flexible reproduction strategies, large population sizes and high standing genetic diversity, traits that should facilitate rapid evolution under directional selection. We quantified local adaptation of copper tolerance in a population of the diatom Skeletonema marinoi from a mining‐exposed inlet in the Baltic Sea and in a non‐exposed population 100 km away. We hypothesized that mining pollution has driven evolution of elevated copper tolerance in the impacted population of S. marinoi. Assays of 58 strains originating from sediment resting stages revealed no difference in the average tolerance to copper between the two populations. However, variation within populations was greater at the mining site, with three strains displaying hyper‐tolerant phenotypes. In an artificial evolution experiment, we used a novel intraspecific metabarcoding locus to track selection and quantify fitness of all 58 strains during co‐cultivation in one control and one toxic copper treatment. As expected, the hyper‐tolerant strains enabled rapid evolution of copper tolerance in the mining‐exposed population through selection on available strain diversity. Within 42 days, in each experimental replicate a single strain dominated (30%–99% abundance) but different strains dominated the different treatments. The reference population developed tolerance beyond expectations primarily due to slowly developing plastic response in one strain, suggesting that different modes of copper tolerance are present in the two populations. Our findings provide novel empirical evidence that standing genetic diversity of phytoplankton resting stage allows populations to evolve rapidly (20–50 generations) and flexibly on timescales relevant for seasonal bloom progressions. [ABSTRACT FROM AUTHOR]

Simple Summary: This first comprehensive genomic analysis of rapidly growing non-tuberculous mycobacteria (RGM) isolates from Betta splendens has led to the re-identification of certain isolates and the discovery of a potential new species, Mycobacterium mucogenicum subsp. phocaicum sp. nov. The in silico analysis is consistent with and confirms previously reported in vitro studies on virulence and antibiotic susceptibility profiles. This study emphasizes the crucial role of whole genome sequencing in the accurate identification and classification of mycobacterial species. This study aims to genomically elucidate six isolates of rapidly growing non-tuberculous mycobacteria (RGM) derived from Siamese fighting fish (Betta splendens). These isolates had previously undergone phenotypic and biochemical characterization, antibiotic susceptibility testing, and in vivo virulence assessment. Initial DNA barcoding using the 16S rRNA sequence assigned these six isolates to five different species, namely Mycobacterium chelonae (BN1983), M. cosmeticum (BN1984 and N041), M. farcinogenes (SNSK5), M. mucogenicum (BN1956), and M. senegalense (BN1985). However, the identification relied solely on the highest percent identity of the 16S rRNA gene, raising concerns about the taxonomic ambiguity of these species. Comprehensive whole genome sequencing (WGS) and extended genomic comparisons using multilocus sequence typing (MLST), average nucleotide identity (ANI), and digital DNA–DNA hybridization (dDDH) led to the reclassification of BN1985 and SNSK5 as M. conceptionense while confirming BN1983 as M. chelonae and BN1984 and N041 as M. cosmeticum. Notably, the analysis of the BN1956 isolate revealed a potential new species that is proposed here as M. mucogenicum subsp. phocaicum sp. nov. Common genes encoding "mycobacterial" virulence proteins, such as PE and PPE family proteins, MCE, and YrbE proteins, were detected in all six isolates. Two species, namely M. chelonae and M. cosmeticum, appear to have horizontally acquired T6SS-II (clpB), catalase (katA), GroEL (groel), and capsule (rmlb) from distantly related environmental bacteria such as Klebsiella sp., Neisseria sp., Clostridium sp., and Streptococcus sp. This study provides the first draft genome sequence of RGM isolates currently circulating in B. splendens and underscores the necessity of WGS for the identification and classification of mycobacterial species. [ABSTRACT FROM AUTHOR]

Spoilage of postharvest strawberries by fungal pathogens is a major global concern. This study employed culture‐dependent and culture‐independent methods to identify postharvest fungal communities of strawberries in Western Australia (WA). Ripe strawberries from eight varieties were sampled twice, early and late in the growing season, from 20 farms in two growing regions. At the first sampling time, traditional isolation and identification demonstrated a high abundance of Botrytis and Cladosporium in Region 1. Mucor was abundant at the second sampling time in Region 1 and both times in Region 2. Metabarcoding confirmed variety‐specific fungal communities in WA strawberries. Botrytis and Cladosporium were predominant in Region 1 and Region 2 samples with more than 50% read abundance except on variety Suncoast. No Mucor sequences were detected using metabarcoding, although they were isolated from all samples using culture‐based methods. Sisquoc, a hydroponic variety, had a significantly higher fungal diversity than the soil‐grown varieties. Varieties Suncoast and Albion exhibited the lowest fungal diversity compared to others. This study demonstrated the value of using culture‐dependent and culture‐independent methods to investigate the complexity of fungal microbiomes of postharvest strawberries. [ABSTRACT FROM AUTHOR]

Describing living community compositions is essential to monitor ecosystems in a rapidly changing world, but it is challenging to produce fast and accurate depiction of ecosystems due to methodological limitations. Morphological methods provide absolute abundances with limited throughput, whereas metabarcoding provides relative abundances of genes that may not correctly represent living communities from environmental DNA assessed with morphological methods. However, it has the potential to deliver fast descriptions of living communities provided that it is interpreted with validated species‐specific calibrations and reference databases. Here, we developed a quantitative approach to retrieve from metabarcoding data the assemblages of living large benthic foraminifera (LBF), photosymbiotic calcifying protists, from Indonesian coral reefs that are under increasing anthropogenic pressure. To depict the diversity, we calculated taxon‐specific correction factors to reduce biological biases by comparing surface area, biovolume and calcite volume, and the number of mitochondrial gene copies in seven common LBF species. To validate the approach, we compared calibrated datasets of morphological communities from mock samples with bulk reef sediment; both sample types were metabarcoded. The calibration of the data significantly improved the estimations of genus relative abundance, with a difference of ±5% on average, allowing for comparison of past morphological datasets with future molecular ones. Our results also highlight the application of our quantitative approach to support reef monitoring operations by capturing fine‐scale processes, such as seasonal and pollution‐driven dynamics, that require high‐throughput sampling treatment. [ABSTRACT FROM AUTHOR]

Molecular techniques like metabarcoding, while promising for exploring diversity of communities, are often impeded by the lack of reference DNA sequences available for taxonomic annotation. Our study explores the benefits of combining targeted DNA barcoding and morphological taxonomy to improve metabarcoding efficiency, using beach meiofauna as a case study. Beaches are globally important ecosystems and are inhabited by meiofauna, microscopic animals living in the interstitial space between the sand grains, which play a key role in coastal biodiversity and ecosystem dynamics. However, research on meiofauna faces challenges due to limited taxonomic expertise and sparse sampling. We generated 775 new cytochrome c oxidase I DNA barcodes from meiofauna specimens collected along the Netherlands' west coast and combined them with the NCBI GenBank database. We analysed alpha and beta diversity in 561 metabarcoding samples from 24 North Sea beaches, a region extensively studied for meiofauna, using both the enriched reference database and the NCBI database without the additional reference barcodes. Our results show a 2.5‐fold increase in sequence annotation and a doubling of species‐level Operational Taxonomic Units (OTUs) identification when annotating the metabarcoding data with the enhanced database. Additionally, our analyses revealed a bell‐shaped curve of OTU richness across the intertidal zone, aligning more closely with morphological analysis patterns, and more defined community dissimilarity patterns between supralittoral and intertidal sites. Our research highlights the importance of expanding molecular reference databases and combining morphological taxonomy with molecular techniques for biodiversity assessments, ultimately improving our understanding of coastal ecosystems. [ABSTRACT FROM AUTHOR]

Sciadonus alphacrucis Melo, Gomes, Møller & Nielsen, 2022 is a rare deep‐sea species, previously known from only two specimens collected off São Paulo State, southeastern Brazil, in the western South Atlantic. Herein, we report a new specimen of S. alphacrucis collected on the continental slope off Santa Catarina State, southern Brazil, thereby extending its known distribution by 420 km. Additionally, we provide the new meristic and morphometric data, the molecular identification using sequences of the cytochrome c oxidase subunit I (COI), an updated distribution map, and a discussion of troglomorphic traits. [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]

Barathronus is a genus of blind cusk eels comprising 11 valid species. In this paper, we report the second specimen ever documented of Barathronus roulei (Bythitidae) obtained from the Porcupine Bank by R.V. Vizconde de Eza using a bottom trawl at a depth of 1349 m. Morphological description and illustrations, including a radiograph, are provided. In addition, three new sequences corresponding to three different genes, cytochrome c oxidase subunit I (COI)‐DNA barcoding, 16S ribosomal RNA (16S), and recombination activating protein 1 (RAG1), have been added to the molecular repositories, representing the first sequences for the species. [ABSTRACT FROM AUTHOR]

The fruit known as wax apple, scientifically named Syzygium samarangense (Blume) Merr. & L.M. Perry, is a well- liked agricultural product originating from Demak Regency in the Central Java Province. When it is difficult to distinguish between different forms of an organism, using DNA barcoding to authenticate the identity of species based on their DNA is an important way. The psbA-trnH intergenic spacer is a commonly used molecular approach to analyse the genetic characteristics of plant species. This study investigated the psbA-trnH intergenic spacer profile of wax apple cultivars from Demak regency. The investigation revealed that the psbA-trnH intergenic spacer sequences of the S. samarangense cultivars, namely Madu Thailand and Madu Deli Hijau, had lengths of 535 and 492 base pairs, respectively. Both cultivars demonstrate a greater nucleotide composition of deoxyadenylic acid (A) and deoxythymidylic acid (T) in comparison to deoxycytidilic acid (C) and deoxyguanylic (G). The genetic distance between S. samarangense ‘Madu Thailand’ and ‘Madu Deli Hijau’ indicates a very tight relationship, with a value of 0.000. The psbA-trnH intergenic spacer proved to be insufficient in differentiating the S. samarangense cultivars from Demak regency, mostly because to its low capacity to discern between the wax apple cultivars. [ABSTRACT FROM AUTHOR]

DNA barcoding is an effective identification tool for tree ferns with heteromorphic generations with morphologically simple gametophytic phase. A complete reference dataset which includes coverage of the target local flora is necessary for accurate identification. Three different Cyathea sp.pl. were investigated to test the utility of plastid DNA barcode regions rbcL with the intention of developing an identification system for native pteridophytes. In addition, the inter-specific and inter-generic variations among the Cyathea sp.pl. were also focused. BLAST analysis revealed the similar rbcL sequences related to C. nilgirensis Holttum, C. gigantea (Wallich ex Hook. f.) Holttum and C. crinita (Hook.) Copel and a sum of 143 sequences viz., Cyathea sp.pl. (121), Gymnosphaera sp.pl.(4), Sphaeropteris sp.pl. (1) and Alsophila sp.pl. (17). The sequences of 143 plants were retrieved from GenBank in FASTA format. The sequences were aligned using multiple sequence alignment tool MUSCLE. The inter-specific and inter-generic distance percentage were calculated using Maximum Likelihood, Minimum Evolution, UPGMA (Unweighted Pair Group Method with Arithmetic Mean) and Neighbor-Joining method. The analyses were constructed using MEGA X software. The rbcL gene provided sequence variation with its strong resolving power. This study demonstrates the overall effectiveness of DNA barcodes for species identification of tree fern genus Cyathea in the pteridophyte flora of Southern Western Ghats. [ABSTRACT FROM AUTHOR]

DNA barcoding is a popular approach that can aid in identifying species and has led to the discovery of many new amphibian species. In this study, we first performed DNA barcoding for two Hylidae family representatives (Hyla orientalis and Hyla savignyi) in the Anatolia region of Türkiye. Five species delineation tests (ABGD, GMYC, PTP, RESL, and statistical parsimony analysis) were applied, and four of them indicated that three mitochondrial lineages of Hylidae are present in Anatolia. Intraspecific genetic distances (K2P) ranged from 0.0014 to 0.0045, while the interspecific genetic distances ranged from 0.0753 to 0.1933. According to the tree topologies obtained from maximum likelihood (ML) within H. savignyi from Anatolia, two lineages can be differentiated, and further investigation is warranted as to whether these lineages represent distinct species. [ABSTRACT FROM AUTHOR]

Thai species of the pill millipede genus Hyleoglomeris Verhoeff, 1910 are reviewed, and an annotated catalogue is provided. Six new species from Thailand are described and illustrated based on morphological characteristics and molecular analyses: H. dracosphaera sp. n., H. nigromaculata sp. n., H. suwannakhuhensis sp. n., H. bomba sp. n., H. tongkerdae sp. n., and H. krasoon sp. n. They are morphologically similar, yet all can be separated based on their characteristic colour patterns, telopod structures and geographic distributions. Genetic differences in the DNA barcode region confirm their full species statuses, with interspecific COI p-distances ranging from 9.01% to 16.51% dissimilarity. We also propose the following new synonym: H. hongkhraiensis Likhitrakarn, Golovatch & Panha, 2015 = H. cavicola Likhitrakarn, Golovatch & Panha, 2015 syn. n. This is based on the low genetic divergence observed between these two taxa (2.34% COI p-distance). A new distribution map of and a key to all Hyleoglomeris species presently known to occur in Thailand are also provided. [ABSTRACT FROM AUTHOR]

Female solitary bees collect pollen for their brood at the expense of using sugars in the nectar as fuel for flight. When flowers provide both pollen and nectar, solitary bees can refuel from the flowers to sustain pollen foraging. However, not all flowers provide both pollen and nectar. This study investigates how females of the solitary bee Andrena taraxaci orienticola secure fuel to sustain pollen foraging when flowers provide scarce nectar. By using DNA barcoding, we identified the Japanese dandelion Taraxacum platycarpum as their primary food source. This dandelion produces almost no nectar in the early morning when female bees collect almost no nectar but collect substantial amounts of pollen. We also found that these bees leave the nest with significantly more nectar at the first flight of the day than at later flights. When we restricted their fuel load by preventing nectar feeding, significantly more females failed to collect pollen on the first flight of the following day. These results suggest females hold back some part of the nectar collected at the end of a day's foraging and use it the following morning as fuel to sustain pollen foraging from flowers providing little nectar. [ABSTRACT FROM AUTHOR]

North American river otters (Lontra canadensis) are semiaquatic mammals distributed throughout the North American continent. They are generalist carnivores with a high potential to affect community structure and dynamics. However, habitat degradation and ecosystem changes have the potential to impact their trophic interactions. Therefore, investigating their diet is critical to understand how anthropogenic disturbances can affect their ecological roles, particularly in the urbanised ecosystems in Florida, where such diet information is currently unavailable. Due to their elusive nature, direct observation of feeding events is a major challenge. Here, we have applied DNA metabarcoding of faecal matter (fDNA), opportunistically collected from river otters killed by road traffic in eastern Florida. Two mitochondrial markers were used to identify prey species' DNA—one for fish and one for vertebrates. A total of 23 unique exact sequence variants (ESVs) were identified, the majority belonging not only to teleosts but also to amphibians, reptiles, birds and mammals. The prey species identified are known dietary resources for river otters and the recovered dietary profiles provided independently by the fish and vertebrate assays were similar. Among the identified teleost prey, three fish species are considered invasive. Here, we show that when sufficient reference sequences are available, fDNA metabarcoding provides high prey species resolution. The collection and metabarcoding of faecal samples is particularly advantageous for scaling diet studies, both geographically and longitudinally, highlighting the potential of fDNA metabarcoding for the investigation of the diet of predators. This can provide foundational information on the organisation of ecological networks, which is vital for implementing successful ecosystem‐based conservation management. [ABSTRACT FROM AUTHOR]

Neogoniolithon is a widespread genus of non‐geniculate coralline algae in the Mediterranean Sea. Three vaguely differentiated species have been reported in the region. This study examined the diversity of Neogoniolithon in the Mediterranean by inferring phylogenies using psbA and COI‐5P markers, applying delimitation algorithms and comparing morpho‐anatomies of putative species. In contrast to previous reports, 13 species were delimited from the Mediterranean in two distant clades. Initial observations suggested that tetrasporangial conceptacle size might be used to separate the two clades. Only three species showed fruticose morphologies. Further morpho‐anatomical differentiation was difficult. One species included Mediterranean, Pacific and Atlantic samples, while three other species occurred in the Mediterranean and the Canary Islands. The global tree revealed that most species were geographically restricted. Crypticity in multiple lineages implied a much higher diversity than currently reported, even lacking samples from the eastern Mediterranean. Only sequence data can be used to identify these cryptic lineages. High endemism emphasizes the importance of conservation of coralline algal bioconstructions to prevent extinctions. The presence of cryptic diversity also invites a reassessment of the knowledge on species distributions that are important in conservation planning and management. [ABSTRACT FROM AUTHOR]

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DNA metabarcoding is increasingly used in dietary studies, but this molecular technique has limitations including DNA extraction, primer choice suitability and completeness of DNA reference databases. Together, these limitations can create successive biases; some of which can be identified by careful data interrogation. Here, we examine some of the influential factors impacting DNA metabarcoding by focusing on the diet of the grazing coral reef parrotfish Scarus rivulatus (Labridae). The 16S V3-4 and 18S V4 gene regions were targeted to detect a wide range of taxa in the pharyngeal content, but genus-level amplicon sequence variant (ASV) assignments were ≤ 40%. Multiple universal 18S V4 primer sets, which amplified the same portion of the gene region, highlighted disparities that impacted resulting datasets and dietary data interpretation. Approximately 50% of ASVs overlapped among these 18S V4 datasets. Overall, our results show that protein-rich epilithic and endolithic microscopic photoautotrophs were consistently present and that S. rivulatus appears to rely on protein-rich taxa, especially dinoflagellates and cyanobacteria. Even if all qualitative limitations could be addressed, our data emphasise the difficulty of making straightforward quantitative conclusions about the diet composition of grazing fishes using DNA metabarcoding alone. Thus, until DNA metabarcoding reaches a state of maturity where quantitative data can be gained, additional support is required from other dietary analysis methods to provide robust interpretations and conclusions. [ABSTRACT FROM AUTHOR]

Detailed knowledge of dietary preferences in wild species are important both for understanding evolutionary processes as well as for their conservation. By means of DNA metabarcoding analysis of faecal samples, we investigated the diet of Alpine ibex (Capra ibex), a mountain ungulate characterised by strong sexual dimorphism and segregation, and analysed differences between male and female diet. Despite a low level of selectivity, we found differences in the diet of males and females that consumed different plant in different quantities. Differences were maintained during both spring and summer. Males tended to feed more on plants richer in fibres (i.e., grasses and sedges) and showed stronger seasonal differences while females consumed more digestible plant families (i.e., forbs). Differences in diet between male and female Alpine ibex are compatible with the strong sexual dimorphism and segregation typical of the species and with female reproductive constraints. These results also show that conservation actions have to take into account different ecological needs of males and females in species characterized by sexual dimorphism and segregation. [ABSTRACT FROM AUTHOR]

Copyright of R.A.R.E - Association Roussillonnaise d'Entomologie is the property of Association Roussillonnaise d Entomologie and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Zoology's dark matter comprises hyperdiverse, poorly known taxa that are numerically dominant but largely unstudied, even in temperate regions where charismatic taxa are well understood. Dark taxa are everywhere, but high diversity, abundance, and small size have historically stymied their study. We demonstrate how entomological dark matter can be elucidated using high-throughput DNA barcoding ("megabarcoding"). We reveal the high abundance and diversity of scuttle flies (Diptera: Phoridae) in Sweden using 31,800 specimens from 37 sites across four seasonal periods. We investigate the number of scuttle fly species in Sweden and the environmental factors driving community changes across time and space. Results: Swedish scuttle fly diversity is much higher than previously known, with 549 putative specie) detected, compared to 374 previously recorded species. Hierarchical Modelling of Species Communities reveals that scuttle fly communities are highly structured by latitude and strongly driven by climatic factors. Large dissimilarities between sites and seasons are driven by turnover rather than nestedness. Climate change is predicted to significantly affect the 47% of species that show significant responses to mean annual temperature. Results were robust regardless of whether haplotype diversity or species-proxies were used as response variables. Additionally, species-level models of common taxa adequately predict overall species richness. Conclusions: Understanding the bulk of the diversity around us is imperative during an era of biodiversity change. We show that dark insect taxa can be efficiently characterised and surveyed with megabarcoding. Undersampling of rare taxa and choice of operational taxonomic units do not alter the main ecological inferences, making it an opportune time to tackle zoology's dark matter. [ABSTRACT FROM AUTHOR]

Dust deposition can supply nutrients to the upper ocean, and subsequently affect primary production and biodiversity in planktonic ecosystem, but the differential response among taxa and their interactions are not fully understood. Here, we performed 7-day onboard incubation experiment amended with different dust loadings (0, 0.2 and 2 mg L-1) in the Kuroshio-Oyashio transition region of the Northwest Pacific Ocean and characterized the community structure and microbial network of a planktonic microbiome in response to dust addition. Chlorophyll a and nutrient analysis indicated that dust-derived nitrogen promoted the growth of phytoplankton (165.8-293.6%) and phytoplankton size structure shifted towards larger cells (>3 µm).Metabarcoding sequencing, targeting prokaryotic and eukaryotic microbes, revealed the changes in community structure and co-occurrence network in response to dust addition. Dust induced a shift from dinoflagellate dominance toward diatom dominance in phytoplankton, and favored members of Cercozoa, Labyrinthulomycetes and Saprospirae, which showed positive correlation with diatom. Temporal response patterns among taxa were categorized into five clusters, and collectively pointed to a more sensitive microeukaryotic community than prokaryotic one in response to dust. The community turnover during the incubation was dominated by moderate taxa with 55.71-62.26% moderate OTUs transitioning to abundant or rare taxa, and dust addition stimulated the transitions of rare taxa. Moreover, biotic factors shaped planktonic microbiome more than abiotic factors, particularly the cross-domain interaction significantly affected microeukaryotic community. Notably, dust addition enhanced the co-occurrence network complexity, with the number of keystone taxa increased, suggesting more interspecies interactions were induced by dust. With integrated analysis, our findings highlight the differential sensitivity of planktonic microbiome to dust deposition and the effects could pass on other organisms through interspecies interaction. [ABSTRACT FROM AUTHOR]

Endemic fish species have long supported the livelihoods of local communities in the Rufiji River Basin (RRB). However, destructive fishing practices have led to a concerning decline in endemic fish stocks. To assess these changes, this study employed key informant interviews, focus group discussions (FGDs), and fishery surveys to assess the historical and contemporary distribution of endemic fishes within the RRB. DNA barcoding was also used to verify species identities. Out of 37 reported fish species, 33 species (54.55% endemic and 45.45% exotic to RRB) were confirmed through DNA barcoding and morphological characteristics. About 5 species including, Heterobranchus longifilis, Citharinus congicus, Labeo congoro, Mormyrus longirostris, and Labeobarbus leleupanus were rarely found in the field, despite being classified as Least Concern by IUCN. Additionally, five species that were reported to be present in the RRB by experienced fishers were not captured during sampling. This highlights the need for validation of the existence of such species through eDNA metabarcoding. Moreover, due to the rarity of some species in the area, their IUCN assessment should be revisited. [ABSTRACT FROM AUTHOR]

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]

Simple Summary: Heterakis spumosa Schneider, 1886 is a parasite commonly found in rodents, primarily associated with the black rat Rattus rattus and the brown rat R. norvegicus, two global invasive rodent species, and the house mouse Mus musculus. This parasite originated probably from Asia, and has been reported in various rodent species, but molecular information from its putative native region is still lacking. Our sampling efforts across Southeast Asia allowed us to collect and analyze this species from two localities in Lao PDR and five localities in Thailand. Additionally, specimens from Europe were included in the analysis. All specimens were analyzed using mtCOI gene and nuclear ribosomal ITS1. Our findings revealed the presence of two distinct clades of H. spumosa, with no discernible association with hosts and geographical localities. Heterakis spumosa, a parasitic worm infecting rodents, is globally prevalent in black rats, brown rats, and house mice. It is hypothesized to originate from Asia due to its widespread presence in Southeast Asia in various Murinae. Previous molecular studies focused on European, African, and Japanese specimens, but none included samples from the putative native range. Rodents were collected between 2008 and 2015 across various localities in Southeast Asia and Europe, identified by morphology or genetic barcoding. Viscera were examined or preserved for later inspection. DNA was extracted from H. spumosa. PCR amplification targeting the mtCOI gene and ITS1 region was conducted in this study using newly designed primers (based on Heterakis reference sequences). PCR amplicons were subsequently sequenced and analyzed. In this study, the phylogenetic analysis using ITS1 sequences revealed that Heterakis samples from Thai and Laotian rodents belong to the species H. spumosa, exhibiting low genetic variation compared to samples from other regions. Genetic distance calculations using mtCOI sequences confirmed the marked distinction of H. spumosa from other Heterakis species. Our phylogenetic analyses using partial mtCOI and ITS1 sequences have significantly enhanced our comprehension of the genetic diversity and evolutionary history of the nematode H. spumosa. [ABSTRACT FROM AUTHOR]

Genome skimming is a novel approach that enables obtaining large-scale genomic information based on high-copy DNA fractions from shallow whole-genome sequencing. The simplicity of this method, low analysis costs, and large amounts of generated data have made it widely used in plant research, including species identification, especially in the case of protected or endangered taxa. This task is particularly difficult in the case of closely related taxa. The Pinus mugo complex includes several dozen closely related taxa occurring in the most important mountain ranges in Europe. The taxonomic rank, origin, or distribution of many of these taxa have been debated for years. In this study, we used genome skimming and multilocus DNA barcoding approaches to obtain different sequence data sets and also to determine their genetic diversity and suitability for distinguishing closely related taxa in the Pinus mugo complex. We generated seven different data sets, which were then analyzed using three discrimination methods, i.e., tree based, distance based, and assembling species by automatic partitioning. Genetic diversity among populations and taxa was also investigated using haplotype network analysis and principal coordinate analysis. The proposed data set based on divergence hotspots is even twenty-times more variable than the other analyzed sets and improves the phylogenetic resolution of the Pinus mugo complex. In light of the obtained results, Pinus × rhaetica does not belong to the Pinus mugo complex and should not be identified with either Pinus uliginosa or Pinus rotundata. It seems to represent a fixed hybrid or introgressant between Pinus sylvestris and Pinus mugo. In turn, Pinus mugo and Pinus uncinata apparently played an important role in the origins of Pinus uliginosa and Pinus rotundata. [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]

Marine sediments host heterogeneous protist communities consisting of both living benthic microorganisms and planktonic resting stages. Despite their key functions in marine ecosystem processes and biogeochemical cycles, their structure and dynamics are largely unknown. In the present study, with a spatially intensive sampling design we investigated benthic protist diversity and function of surface sediment samples from three subregions of the Mediterranean Sea, through an environmental DNA metabarcoding approach targeting the 18S V4 region of rRNA gene. Protists were characterized at the taxonomic level and trophic function, both in terms of alpha diversity and community composition, testing for potential differences among marine subregions and bathymetric groups. Overall, Alveolata and Stramenopiles were the two divisions that dominated the communities. These dominant groups exhibited significant differences among the three Mediterranean subregions in the alpha diversity estimates based on the detected ASVs, for all computed indices (ASV richness, Shannon and Simpson indices). Protist communities were also found to be significantly different in terms of composition at the order rank in the three subregions p-value < 0.01). These differences were mainly driven by Anoecales, Peridiniales, Borokales, Paraliales and Gonyaulacales, which together contributed almost 80% of the average dissimilarity. Anoecales was the dominant order in the Ionian – Central Mediterranean and Adriatic Sea, but with considerably different relative abundances (52% and 36%, respectively), while Borokales was the dominant order in the Western Mediterranean Sea (33%). Similarly, significant differences among the three marine subregions were also highlighted when protist assemblages were examined in terms of trophic function, both in terms of alpha diversity (calculated on the ASVs for each trophic group) and community composition p-value < 0.01. In particular, the Adriatic Sea stood out for having the highest relative abundance of autotrophic/ mixotrophic components in the surface sediments analyzed. Conversely, no significant differences in protist assemblages were found among depth groups. This study provided new insights into the taxonomic and trophic composition of benthic protist communities found in Mediterranean surface sediments, revealing geographical differences among regional seas. The results were discussed in relation to the Mediterranean environmental features that could generate the differences among benthic protist communities. [ABSTRACT FROM AUTHOR]

Simarouba, a neotropical genus in the family Simaroubaceae, currently lacks comprehensive genomic data in existing databases. This study aims to fill this gap by providing genomic resources for three Simarouba species, S. amara, S. versicolor, and S. glauca. It also aims to perform comparative molecular evolutionary analyses in relation to other species within the order Sapindales. The analysis of these three Simarouba species revealed the presence of the typical quadripartite structure expected in plastomes. However, some pseudogenization events were identified in the psbC, infA, rpl22, and ycf1 genes. In particular, the CDS of the psbC gene in S. amara was reduced from 1422 bp to 584 bp due to a premature stop codon. Nucleotide diversity data pointed to gene and intergenic regions as promising candidates for species and family discrimination within the group, specifically matK, ycf1, ndhF, rpl32, petA-psbJ, and trnS-trnG. Selection signal analyses showed strong evidence for positive selection on the rpl23 gene. Phylogenetic analyses indicated that S. versicolor and S. glauca have a closer phylogenetic relationship than S. amara. We provide chloroplast genomes of three Simaruba species and use them to elucidate plastome evolution, highlight the presence of pseudogenization, and identify potential DNA barcode regions. [ABSTRACT FROM AUTHOR]

Background: Western Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India. Methods: A comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis. Results: Phlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species. Conclusions: The Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly. [ABSTRACT FROM AUTHOR]

Background: DNA metabarcoding applies high-throughput sequencing approaches to generate numerous DNA barcodes from mixed sample pools for mass species identification and community characterisation. To date, however, most metabarcoding studies employ second-generation sequencing platforms like Illumina, which are limited by short read lengths and longer turnaround times. While third-generation platforms such as the MinION (Oxford Nanopore Technologies) can sequence longer reads and even in real-time, application of these platforms for metabarcoding has remained limited possibly due to the relatively high read error rates as well as the paucity of specialised software for processing such reads. Results: We show that this is no longer the case by performing nanopore-based, cytochrome c oxidase subunit I (COI) metabarcoding on 34 zooplankton bulk samples, and benchmarking the results against conventional Illumina MiSeq sequencing. Nanopore R10.3 sequencing chemistry and super accurate (SUP) basecalling model reduced raw read error rates to ~ 4%, and consensus calling with amplicon_sorter (without further error correction) generated metabarcodes that were ≤ 1% erroneous. Although Illumina recovered a higher number of molecular operational taxonomic units (MOTUs) than nanopore sequencing (589 vs. 471), we found no significant differences in the zooplankton communities inferred between the sequencing platforms. Importantly, 406 of 444 (91.4%) shared MOTUs between Illumina and nanopore were also found to be free of indel errors, and 85% of the zooplankton richness could be recovered after just 12–15 h of sequencing. Conclusion: Our results demonstrate that nanopore sequencing can generate metabarcodes with Illumina-like accuracy, and we are the first study to show that nanopore metabarcodes are almost always indel-free. We also show that nanopore metabarcoding is viable for characterising species-rich communities rapidly, and that the same ecological conclusions can be obtained regardless of the sequencing platform used. Collectively, our study inspires confidence in nanopore sequencing and paves the way for greater utilisation of nanopore technology in various metabarcoding applications. [ABSTRACT FROM AUTHOR]

Environmental DNA (eDNA) is becoming an increasingly important tool in diverse scientific fields from ecological biomonitoring to wastewater surveillance of viruses. The fundamental challenge in eDNA analyses has been the bioinformatical assignment of reads to taxonomic groups. It has long been known that full probabilistic methods for phylogenetic assignment are preferable, but unfortunately, such methods are computationally intensive and are typically inapplicable to modern next-generation sequencing data. We present a fast approximate likelihood method for phylogenetic assignment of DNA sequences. Applying the new method to several mock communities and simulated datasets, we show that it identifies more reads at both high and low taxonomic levels more accurately than other leading methods. The advantage of the method is particularly apparent in the presence of polymorphisms and/or sequencing errors and when the true species is not represented in the reference database. [ABSTRACT FROM AUTHOR]

Most host-parasite associations are explained by phylogenetically conservative capabilities for host utilization, and therefore parasite switches between distantly related hosts are rare. Here we report the first evidence of a parasitic spillover of the burrowing sea anemone Edwardsiella carnea from the invasive ctenophore Mnemiopsis leidyi to two scyphozoan hosts: the native Mediterranean barrel jellyfish Rhizostoma pulmo and the invasive Indo-Pacific nomad jellyfish Rhopilema nomadica, collected from the Eastern Mediterranean Sea. Edwardsiella carnea planulae found in these jellyfish were identified using molecular analyses of the mitochondrial 16S and nuclear 18S rRNA genes. Overall, 93 planulae were found on tentacles, oral arms, and inside of the gastrovascular canals of the scyphomedusae, whereas no infection was observed in co-occurring ctenophores. DNA metabarcoding approach indicated seasonal presence of Edwardsiella sp. in the Eastern Mediterranean mesozooplankton, coinciding with jellyfish blooms in the region. Our findings suggest a non-specific parasitic relationship between Edwardsiella carnea and various gelatinous hosts based on shared functionality rather than evolutionary history, potentially driven by shifts in host availability due to jellyfish blooms. This spillover raises questions about the ecological impacts of parasitism on native and invasive scyphozoan hosts and the potential role of Edwardsiella in controlling their populations. [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]

Improved understanding of mosquito–plant feeding interactions can reveal insights into the ecological dynamics of pathogen transmission. In wild malaria vectors Anopheles gambiae s.l. and An. funestus group surveyed in selected dryland ecosystems of Kenya, we found a low level of plant feeding (2.8%) using biochemical cold anthrone test but uncovered 14-fold (41%) higher rate via DNA barcoding targeting the chloroplast rbcL gene. Plasmodium falciparum positivity was associated with either reduced or increased total sugar levels and varied by mosquito species. Gut analysis revealed the mosquitoes to frequently feed on acacia plants (~ 89%) (mainly Vachellia tortilis) in the family Fabaceae. Chemical analysis revealed 1-octen-3-ol (29.9%) as the dominant mosquito attractant, and the sugars glucose, sucrose, fructose, talose and inositol enriched in the vegetative parts, of acacia plants. Nutritional analysis of An. longipalpis C with high plant feeding rates detected fewer sugars (glucose, talose, fructose) compared to acacia plants. These results demonstrate (i) the sensitivity of DNA barcoding to detect plant feeding in malaria vectors, (ii) Plasmodium infection status affects energetic reserves of wild anopheline vectors and (iii) nutrient content and olfactory cues likely represent potent correlates of acacia preferred as a host plant by diverse malaria vectors. The results have relevance in the development of odor-bait control strategies including attractive targeted sugar-baits. [ABSTRACT FROM AUTHOR]

Identification of fish larvae based on morphology is typically limited to higher taxonomic ranks (e.g., family or order), as larvae possess few morphological diagnostic characters for precise discrimination to species. When many samples are presented at any one time, the use of morphology to identify such specimens can be laborious and time‐consuming. Using a reverse workflow for specimen sorting and identification leveraging high‐throughput DNA sequencing, thousands of fish larvae can be DNA barcoded and sorted into molecular operational taxonomic units (mOTUs) in a single sequencing run with the nanopore sequencing technology (e.g., MinION). This process reduces the time and financial costs of morphology‐based sorting and instead deploys experienced taxonomists for species taxonomic work where they are needed most. In this study, a total of 3022 fish larval specimens from plankton tows across four sites in Singapore were collected and sorted based on this workflow. Eye tissue from individual samples was used for DNA extraction and sequencing of cytochrome c oxidase subunit I. We generated a total of 2746 barcodes after quality filtering (90.9% barcoding success), identified 2067 DNA barcodes (75.3% identification success), and delimited 256 mOTUs (146 genera, 52 families). Our analyses identified specific challenges to species assignment, such as the potential misidentification of publicly available sequences used as reference barcodes. We highlighted how the conservative application and comparison of a local sequence database can help resolve identification conflicts. Overall, this proposed approach enables and expedites taxonomic identification of fish larvae, contributing to the enhancement of reference barcode databases and potentially better understanding of fish connectivity. [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 genus Dicellophilus Cook, 1896, is a peculiar genus from the point of view of distribution. Dicellophilus is distributed in three limited areas that are well separated from one another: central Europe (D. carniolensis), Honshu (D. pulcher), and the southwestern part of the USA (D. anomalus and D. limatus). In the present study, in a field survey conducted throughout Japan, specimens belonging to the genus Dicellophilus were collected from Tohoku to the Kansai region, Honshu. Morphological analysis, molecular phylogenetic analysis, and genetic distance among Dicellophilus in Japan and D. carniolensis revealed that specimens from Sendai-shi, Miyagi Pref., could be assigned to an undescribed species. This previously unrecognized species is herein described as D. praetermissus sp. nov. The new species can be distinguished from D. carniolensis and D. limatus by the number of pairs of legs (43 pairs in D. carniolensis and 45 in D. limatus, but 41 in D. praetermissus sp. nov.), from D. anomalus by the lack of a pair of setae on the posteromedian part of the clypeus and variable crenulation on the internal margin of the forcipular tarsungulum, and from D. pulcher based on the following combination of characteristics: both ends of the transverse suture not evidently convex forward; long rather than wide trochanteroprefemur; wide rather than long metasternite. [ABSTRACT FROM AUTHOR]

The morphology, phylogenetic position, and distribution of a recently revalidated species of leuciscid minnow, Phoxinus isetensis, were substantially clarified. The species was described in the late 18th century from the Middle Urals but later synonymized with Phoxinus phoxinus. As believed, P. isetensis is distributed in the Arctic Ocean catchment from the Murman coast via West and East Siberia until the Pacific Ocean catchment in Far East and Northeastern Asia. Our study, with the use of mtDNA markers coupled with extensive morphological data, showed that the distribution of P. isetensis is greatly different. Currently, this is the most widespread Phoxinus species in Europe, distributed in Northern and Eastern Europe and on the western edge of Siberia (Iset and Ural basins). In particular, P. isetensis inhabits the basins of the Caspian, Baltic, White, Barents, and Kara seas, possibly occurring in the North Sea basin. The species was redescribed, and the type locality and neotype were designated. The main morphological difference from other Phoxinus spp. is the large total number of vertebrae (39–43, mode 41) due to an increase in the number of caudal vertebrae (16–21, mode 19). Phylogenetically, P. isetensis is a sister to the Caucasian species P. colchicus (p-distance = 5%). The wide distribution of P. isetensis within the area of the Last Glacial Maximum suggests rapid colonization of deglaciated regions, probably due to its adaptation to a cold climate. [ABSTRACT FROM AUTHOR]

Pheretimoid earthworms, Amynthas carnosus, were collected from Northeast and North China. An update on the distribution and systematics of the A. carnosus complex in eastern Asia using both morphological and molecular data is provided. Three subspecies, A. carnosus carnosus, A. carnosus naribunji, and A. carnosus roki, are confirmed. Comparisons of morphological characters between the subspecies of A. carnosus are provided. Our results support the subspecies assignment with an intraspecific K2P genetic distance of not greater than 10% using the mitochondrial cytochrome c oxidase subunit I (COI). In addition, a re -description of the morphology of A. carnosus naribunji is presented here. [ABSTRACT FROM AUTHOR]