Phytoplankton community structure in a subtropical river revealed by DNA metabarcoding, with a focus on cell-cyst coupling of Unruhdinium spp. (Dinophyceae).

The concentrations of six sulfonamides (SAs) and three tetracyclines (TCs) were investigated in Jiulongjiang River during the low water season and the high water season. They were monitored in both surface water and sediment. Total concentrations of all these antibiotics varied from 31 to 25,771 ng g(-1) in sediment samples. In water they ranged from 60 to 2607 ng L(-1) during the low water season and from ND (not detected) to 134 ng L(-1) during the high water season. At the sites nearby breeding farms, chlorotetracycline was found to have the highest concentration of 1036 ng L(-1) in water and 14,666 ng g(-1) in sediments. According to the published data, the concentrations of sulfamethazine, sulfameter and TCs at these sites were higher than that in most rivers. The concentrations during the low water season were tens to hundreds of times higher than that in the high water season. The lower concentrations of TCs in the high water season might result from both dilution and photo-degradation, while dilution and bio-degradation might lead to the lower concentrations of SAs. However, further study is needed to clarify the specific reasons. Concerning the relationship between sediment and water samples, the pseudo-partitioning values of TCs were much higher than SAs. It indicates that the TCs are prone to accumulate in the sediment.

Diversity and distribution of harmful microalgae in the Gulf of Thailand assessed by DNA metabarcoding

BackgroundEutrophication of freshwater systems can result in blooms of phytoplankton, in many cases cyanobacteria. This can lead to shifts in structure and functions of phytoplankton communities adversely affecting the quality of drinking water sources, which in turn impairs public health. Relationships between structures of phytoplankton communities and concentrations of the toxicant, microcystin–leucine–arginine (MC-LR), have not been well examined in large shallow lakes. The present study investigated phytoplankton communities at seven locations from January to December of 2015 in Tai Lake, and relationships between structures and diversities of phytoplankton communities and water quality parameters, including concentrations of MC-LR and metals, were analyzed.ResultsA total of 124 taxa of phytoplankton were observed, and the predominant taxa were Microcystis sp. and Dolichospermum flos-aquae of Cyanophyta and Planctonema sp. of Chlorophyta. The greatest diversities of phytoplankton communities, as indicated by species richness, Simpson, Shannon–Wiener, the Berger and Parker, and the Pielou evenness indices, were observed in spring. Furthermore, productivity of phytoplankton was significantly and negatively correlated with diversities. These results demonstrated that Simpson, Shannon–Wiener, the Berger and Parker, and the Pielou evenness indices of phytoplankton communities were significantly related to trophic status and overall primary productivity in Tai Lake. In addition, temperature of surface water, pH, permanganate index, biochemical oxygen demand, total phosphorus, arsenic, total nitrogen/total phosphorous ratio, and MC-LR were the main factors associated with structures of phytoplankton communities in Tai Lake.ConclusionThe present study provided helpful information on phytoplankton community structure and diversity in Tai Lake from January to December of 2015. Our findings demonstrated that Simpson, Shannon–Wiener, the Berger and Parker, and the Pielou evenness indices could be used to assess and monitor for status and trends in water quality of Tai Lake. In addition, MC-LR was one of the main factors associated with structures of phytoplankton communities in Tai Lake. The findings may help to address important ecological questions about the impact of a changing environment on biodiversity of lake ecosystems and the control of algae bloom. Further studies are needed to explore the relationship between MC-LR and phytoplankton communities in the laboratory.

Phytoplankton phenology and community structure in the western North Pacific were investigated for 2001–2009, based on satellite ocean colour data and the Continuous Plankton Recorder survey. We estimated the timing of the spring bloom based on the cumulative sum satellite chlorophyll adata, and found that the Pacific Decadal Oscillation (PDO)‐related interannual SST anomaly in spring significantly affected phytoplankton phenology. The bloom occurred either later or earlier in years of positive or negative PDO (indicating cold and warm conditions, respectively). Phytoplankton composition in the early summer varied depending on the magnitude of seasonal SST increases, rather than the SST value itself. Interannual variations in diatom abundance and the relative abundance of non‐diatoms were positively correlated with SST increases for March–April and May–July, respectively, suggesting that mixed layer environmental factors, such as light availability and nutrient stoichiometry, determine shifts in phytoplankton community structure. Our study emphasised the importance of the interannual variation in climate‐induced warm–cool cycles as one of the key mechanisms linking climatic forcing and lower trophic level ecosystems.

Urbanization reduces resource use efficiency of phytoplankton community by altering the environment and decreasing biodiversity

Extensive genetic diversity and endemism across the global range of the oceanic copepod Pleuromamma abdominalis

Acremonium-like fungi are emerging as important opportunistic pathogens in cutaneous, subcutaneous and serious invasive infections, especially in immunocompromised and debilitated individuals, and Acremonium infections are usually resistant to antifungal therapy. Several molecular studies have demonstrated that many species in the genus Acremonium are polyphyletic, and currently, the genus is restricted to the family Bionectriaceae (Hypocreales). Molecular identification and in vitro antifungal susceptibility tests of Acremonium-like fungi isolated from human clinical specimens in China were performed in this study. Three genetic loci: the large subunit ribosomal RNA gene (LSU), ribosomal internal transcribed spacer and elongation factor 1-α (EF1-α), were used to assess their taxonomic position for correct identification among various species. The multilocus study of twenty-eight strains showed that these strains were distributed in three main lineages: egyptiacum, Cordycipitaceae and Sarocladium; Acremonium egyptiacum and Sarocladium kiliense were the main species of these strains, and three isolates were too phylogenetically distant to be considered undescribed species. Relatively low minimum inhibitory concentrations (MICs) of 0.25-2 and 0.031-0.5μg/mL were found for voriconazole and terbinafine for most species, respectively. Varied antifungal activities of ciclopirox olamine, amorolfine and posaconazole were found in our study. However, no antifungal effect of sertaconazole, itraconazole or fluconazole was observed against most strains. This is the first study on Acremonium-like species diversity by multilocus sequence analyses and antifungal susceptibility of clinically relevant isolates in China.

The genus Scleroderma (Sclerodermataceae) contains gasteroid ectomycorrhizal fungi and is distributed worldwide in temperate and tropical regions. Fresh specimens were collected in Thailand and report three undescribed species and one new record for the country. These species were characterized by photographs of freshly collected basidiomes, and their macro- and microscopic features were compared with those of known species of Scleroderma. Additionally, DNA sequence data were generated for four loci, including the nuclear ribosomal internal transcribed spacer region (ITS), the large subunit ribosomal RNA gene (LSU), the translation elongation factor 1-alpha gene (tef1-α), and the second largest subunit of RNA polymerase II (rpb2). A multi-locus phylogeny was constructed to confirm their taxonomic placement.

DNA metabarcoding is increasingly used for the assessment of aquatic communities, and numerous studies have investigated the consistency of this technique with traditional morpho‐taxonomic approaches. These individual studies have used DNA metabarcoding to assess diversity and community structure of aquatic organisms both in marine and freshwater systems globally over the last decade. However, a systematic analysis of the comparability and effectiveness of DNA‐based community assessment across all of these studies has hitherto been lacking. Here, we performed the first meta‐analysis of available studies comparing traditional methods and DNA metabarcoding to measure and assess biological diversity of key aquatic groups, including plankton, microphytobentos, macroinvertebrates, and fish. Across 215 data sets, we found that DNA metabarcoding provides richness estimates that are globally consistent to those obtained using traditional methods, both at local and regional scale. DNA metabarcoding also generates species inventories that are highly congruent with traditional methods for fish. Contrastingly, species inventories of plankton, microphytobenthos and macroinvertebrates obtained by DNA metabarcoding showed pronounced differences to traditional methods, missing some taxa but at the same time detecting otherwise overseen diversity. The method is generally sufficiently advanced to study the composition of fish communities and replace more invasive traditional methods. For smaller organisms, like macroinvertebrates, plankton and microphytobenthos, DNA metabarcoding may continue to give complementary rather than identical estimates compared to traditional approaches. Systematic and comparable data collection will increase the understanding of different aspects of this complementarity, and increase the effectiveness of the method and adequate interpretation of the results.

Environmental DNA metabarcoding reveals the effect of environmental selection on phytoplankton community structure along a subtropical river

Adopting the currently used concept for the genus Peltaster, the sooty blotch fungus Peltaster cerophilus is newly described from the cuticle of ripening or ripe apples. It forms a punctate phenotype consisting of superficially formed pycnothyria and a superficial mycelial mat consisting of a net of brown or brownish black hyphae. The pycnothyria are olivaceous brown to brown but have a spot in the center that is less strongly pigmented. Pycnothyria on the holotype of P. fructicola are homogeneously pigmented. On synthetic nutrient-poor agar, P. cerophilus is largely indistinguishable from P. fructicola. It forms delicate, spreading hyphae and intercalary conidiogenous cells with short, lateral, apically thick-walled conidiogenous necks forming blastic, unpigmented, one-celled conidia in basipetal succession. Conidia can swell and become one-septate. The species has microcyclical conidiation in proximate parts of colonies. DNA sequence analyses based on the ITS and the partial nuclear small and large subunit ribosomal RNA genes, the partial mitochondrial small subunit rRNA gene and the partial translation elongation factor 1-α gene support the distinction of the European P. cerophilus from P. fructicola, which is known from North America and Europe. The nuclear small ribosomal RNA subunit gene sequences of P. cerophilus contain two group I introns at locations known to accommodate introns in certain other, unrelated taxa. One of these, for which the code “SSU-1506 intron” was adopted, is 1459 base pairs long and located between the universal primer sites ITS5 and ITS1. Similar or positional-differing introns were encountered also in three currently undescribed Peltaster species. Representative strains of Peltaster fructicola did not accommodate introns in the nuclear small subunit ribosomal RNA gene.

Dwelling in a variety of aquatic habitats, one of the most abundant groups of microcrustaceans, ostracodes, are widely used indicator organisms in paleolimnological research. Typically, they are identified via traditional methods using morphological features but this may be excessively time‐consuming and prone to inter‐investigator variation. DNA barcoding and metabarcoding have become important tools for specimen identification, with a great impact in the field of taxonomy, (paleo‐)ecology and evolution. Despite its potential, metabarcoding has been rarely used to analyze the community structure of ostracodes. Here, we evaluate the performance of a metabarcoding approach for ostracode identification in surface sediment samples collected from Lake Nam Co on the Tibetan Plateau. We tested six different primer pairs amplifying fragments of three different genes, and compared their success in inferring ostracode communities, coupled with morphological identification of ostracodes from the same sediment samples. In total, depending on the primer pair used, seven to nineteen ostracode amplicon sequence variants (ASVs) were identified. Via microscopy, eight morphospecies were identified. We found considerable differences between primer pairs in yielding ostracode sequences via metabarcoding. In general, the highest proportions of ostracode reads and ASVs were found with primers amplifying fragments of the 18S rRNA gene, whereas primers for COI gene had the highest in silico amplification success and highest sequencing depth per sample but only contained <1% of ostracode sequences. As a consequence, the metabarcoding results with 18S rRNA gene were more consistent with the morphological data compared to those obtained with COI or mitochondrial 16S rRNA primers. No significant effects of treatment with different sediment quantities for DNA extraction (10 g vs. 0.5 g) were found on ostracode ASVs community composition. These results indicate that DNA metabarcoding can serve as an efficient tool for ostracode‐based environmental reconstructions but requires an informed decision on primers and target gene, as well as extending the barcoding database for improved accuracy.

ABSTRACTThis study investigates the metazoan biodiversity in the Southern Adriatic Sea using environmental DNA (eDNA) metabarcoding. Sediment and adjacent water samples were collected from three sites (one pristine, two impacted by human activities) at three distances from the coast across two seasons. The complex four‐factor experimental design (576 samples) addresses key sources of eDNA variability and provides a valuable comparison of markers (COI and 18S) and sample types, which remain rare in the literature. Results showed differences in the number and type of taxa identified, taxonomic resolution, and number of amplicon sequence variants (ASV) per operational taxonomic unit (OTU) across markers. The obtained overall community structure (beta‐diversity) was similar for both markers. Sediment samples had higher OTU richness, but lower diversity than water samples. The two sample types provided distinct and only partially overlapping views of biodiversity. Sediment samples were rich in benthic species, whereas water samples featured mostly planktonic and nektonic species. Biodiversity varied by site and season, with sediment samples showing less seasonal variability. The pristine site did not host higher biodiversity than impacted sites, likely because of the latter's habitat heterogeneity. This study confirms the effectiveness of eDNA metabarcoding for biodiversity assessment in coastal ecosystems and provides a foundational dataset for future monitoring. By highlighting the complementary nature of COI and 18S markers and the role of sample type, this research supports integrating eDNA metabarcoding into routine environmental monitoring programs while emphasising the need for further standardisation and improved reference databases.

Co-occurrence of microplastics and heavy metals in a freshwater lake system in Indian Himalaya: Distribution and influencing factors

DNA metabarcoding is widely used to study prokaryotic and eukaryotic microbial diversity. Technological constraints limit most studies to marker lengths below 600base pairs (bp). Longer sequencing reads of several thousand bp are now possible with third-generation sequencing. Increased marker lengths provide greater taxonomic resolution and allow for phylogenetic methods of classification, but longer reads may be subject to higher rates of sequencing error and chimera formation. In addition, most bioinformatics tools for DNA metabarcoding were designed for short reads and are therefore unsuitable. Here, we used Pacific Biosciences circular consensus sequencing (CCS) to DNA-metabarcode environmental samples using a ca. 4,500bp marker that included most of the eukaryote SSU and LSU rRNA genes and the complete ITS region. We developed an analysis pipeline that reduced error rates to levels comparable to short-read platforms. Validation using a mock community indicated that our pipeline detected 98% of chimeras de novo. We recovered 947 OTUs from water and sediment samples from a natural lake, 848 of which could be classified to phylum, 397 to genus and 330 to species. By allowing for the simultaneous use of three databases (Unite, SILVA and RDP LSU), long-read DNA metabarcoding provided better taxonomic resolution than any single marker. We foresee the use of long reads enabling the cross-validation of reference sequences and the synthesis of ribosomal rRNA gene databases. The universal nature of the rRNA operon and our recovery of >100 nonfungal OTUs indicate that long-read DNA metabarcoding holds promise for studies of eukaryotic diversity more broadly.