Diatoms as indicators in running waters: trends of studies on biological assessment and monitoring.

Despite the biodiversity and ecosystem services provided by lotic ecosystems, they are strongly affected by anthropogenic activities. Therefore, biological monitoring and assessment strategies are crucial in helping maintain these ecosystems and developing mitigation policies. We provide a global overview of the use of benthic diatoms as bioindicators in lotic environments, by analyzing 764 articles published in the past 20 years. We analyzed the influence of substrate type on samplings, which species have been highlighted as indicators and for which type of impacts, which anthropogenic impacts have been most commonly evaluated, and which metrics have been commonly used in studies using diatoms to assess and monitor the quality of lotic environments. We found that the most studied anthropogenic impact is artificial eutrophication and that some species, especially Nitzschia palea, have been thoroughly mentioned as indicators of this impact. Indicator species related to other types of impact are less common, demonstrating the need for studies on this issue. Moreover, we verified that traditional taxonomic metrics, such as diversity and diatom indices, have been widely used. Some alternative metrics have been used recently, such as those based on teratological valves, lipid bodies, valve size, and DNA metabarcoding. The number of biomonitoring and assessment studies based on diatoms has increased considerably in the past 20 years. Nonetheless, the demand for natural resources and consequently the degradation of lotic ecosystems have accelerated significantly. Thus, the development of low-cost and time-efficient biological assessment and monitoring strategies is essential for evaluating the health of lotic environments.

Reduction of taxonomic bias in diatom species data.

Inconsistency in taxonomic identification and analyst bias impede the effective use of diatom data in regional and national stream and lake surveys. In this study, we evaluated the effect of existing protocols and a revised protocol on the precision of diatom species counts. The revised protocol adjusts five elements of sample preparation, taxon identification and enumeration, and quality control (QC) samples. We used six independent datasets to assess the effect of the adjustments on analytical outcomes. The first dataset was produced by five analysts from three laboratories following a standard protocol (Charles et al. 2002). The remaining datasets were produced by 2-3 analysts in 1-3 laboratories following a revised protocol. The revised protocol included the following modifications: 1) use of Battarbee settling chambers to prepare coverslips, 2) development of coordinated pre-count voucher floras based on morphological operational taxonomic units (mOTUs), 3) random assignment of samples to analysts, 4) post-count identification and documentation of taxa, and 5) increased QC samples. The revised protocol reduced taxonomic bias, as measured by reduction in analyst signal, and improved similarity among QC samples. Reduced taxonomic bias improves the performance of biological assessments, facilitates transparency across studies, and refines estimates of diatom species distributions.

Life cycle size dynamics in Didymosphenia geminata (Bacillariophyceae).

Didymosphenia geminata has received a great deal of attention in the last 25 years, and considerable effort has gone into determining the origin, ecological impact, and economic consequences of its invasive behavior. While environmental conditions are a controlling influence in distribution, the extreme success of the species may be tied to its basic biology and life history. Little is known, however, about population dynamics, size restoration and reproduction of D. geminata. The objective of this study was to determine the temporal patterns in cell size frequency, size restoration strategy, and synchronization of life cycles between populations in close proximity. We implemented FlowCam technology to measure the length of more than 100,000 D. geminata cells from two sites in South Boulder Creek, Colorado over 1 year. We applied finite mixture modeling to uncover temporal patterns in size distribution. Our results show that collections of D. geminata exhibited a complex, multimodal size distribution, almost always containing four overlapping age cohorts. We failed to observe direct visual evidence of the sexual phase. Multiple abrupt and directional shifts in size distribution, however, were documented providing conclusive evidence of cell size restoration. Lastly, nodules in close proximity were asynchronous with respect to size frequency profiles and size diminution, highlighting the relevance of spatial heterogeneity in in situ diatom size dynamics. This study is the first to document the complexity of diatom cell size distribution in a lotic system, size restoration in D. geminata, and the variability in rates of size reduction at microhabitat spatial scales.

Algal Biodiversity of Nine Megaliths in South-East Bulgaria.

This paper presents the first data on the biodiversity of lithophytic algae from Bulgarian megaliths obtained after the application of the direct sampling method, subsequent cultivation, and processing by light microscopy. A rich algal flora was found: 90 species and 1 variety of 65 genera from Cyanoprokaryota/Cyanobacteria (29 species, 13 genera), Chlorophyta (40 species and 1 variety, 38 genera), Streptophyta (5 species, 1 genus), and Ochrophyta (16 species, 13 genera). Among them were the globally rare Pseudodictyochloris multinucleata (Chlorophyta), found for the first time in such lowland and warm habitats, and Scotiella tuberculata (Chlorophyta), for which this is the first finding in the country. Three of the recorded species are conservationally important. The low floristic similarity between the sites (0-33%) shows the diversity of the algal flora, with no common species found for all the megaliths studied. The most widespread were the strongly adaptive and competitive Stichococcus bacillaris, Apatococcus lobatus, and Chloroidium ellipsoidium (Chlorophyta). The correlations estimated between the species number and substrate temperature (18.1-49.6 °C) suggest the prospect of future research related to the impact of global warming. In addition, the study points to the safety aspects as it revealed species from nine potentially toxin-producing cyanoprokaryotic genera that could be harmful to visitors' health.

The Diatom Diversity and Ecological Status of a Tufa-Depositing River through eDNA Metabarcoding vs. a Morphological Approach-A Case Study of the Una River (Bosnia and Herzegovina).

Tufa deposits in karst rivers are unique habitats created by mutual interactions between specific environmental and biotope features and inhabited by diatoms as a highly abundant and diverse algal group. This pilot study aimed to investigate the diversity of diatom communities on tufa depositing habitats and assess the Una River's ecological status using a comparative molecular and morphological approach for diatom identification. The 312 base pairs of the rbcL gene were barcoded and analyzed using MiSeq reads and amplicon sequence variants (ASVs) obtained by the DADA2 pipeline. The reference database Diat.barcode v7 was used for taxonomic assignment. The morphological identification of the diatoms was carried out in parallel. In total, the combined dataset revealed 46 taxa identified at genus rank, 125 on the subgenus, and 145 on combined taxonomy rank. The metabarcoding approach mostly leads to a lower number of identified taxa at species rank (58 in molecular vs. 119 in optical inventory), resulting in higher values of beta diversity and heterogeneity in diatom assemblages in samples obtained by morphological approach. Despite the high percentage of taxonomically not assigned diatom ASVs to the species rank, high Shannon diversity index values and a similar number of taxa per locations compared to the morphological approach were obtained. Taxa Achnanthidium minutissimum (Kützing) Czarnecki, Achnanthidium pyrenaicum (Hustedt) H.Kobayasi, Amphora pediculus (Kützing) Grunow, Diatoma vulgaris Bory, Navicula cryptotenella Lange-Bertalot, and Navicula tripunctata (O.F.Müller) Bory were identified at all locations in both inventories. Although limited consistency in the diatom abundances between the two inventory datasets was found, a similar grouping of samples was observed connected to the river's longitudinal gradient. The data obtained using molecular approach in most sites indicated a mostly lower ecological status (good or moderate) compared to the data obtained from the morphological approach (high, good, and moderate). The potential of environmental DNA (eDNA) diatom metabarcoding for water monitoring and diversity studies is undeniable, but to fully realize the benefits of these methods in the future, it is essential to standardize protocols and expand the reference database for species found in specific habitats, such as tufa deposits.

Seascape analysis reveals regional gene flow patterns among populations of a marine planktonic diatom.

We investigated the gene flow of the common marine diatom, Skeletonema marinoi, in Scandinavian waters and tested the null hypothesis of panmixia. Sediment samples were collected from the Danish Straits, Kattegat and Skagerrak. Individual strains were established from germinated resting stages. A total of 350 individuals were genotyped by eight microsatellite markers. Conventional F-statistics showed significant differentiation between the samples. We therefore investigated whether the genetic structure could be explained using genetic models based on isolation by distance (IBD) or by oceanographic connectivity. Patterns of oceanographic circulation are seasonally dependent and therefore we estimated how well local oceanographic connectivity explains gene flow month by month. We found no significant relationship between genetic differentiation and geographical distance. Instead, the genetic structure of this dominant marine primary producer is best explained by local oceanographic connectivity promoting gene flow in a primarily south to north direction throughout the year. Oceanographic data were consistent with the significant FST values between several pairs of samples. Because even a small amount of genetic exchange prevents the accumulation of genetic differences in F-statistics, we hypothesize that local retention at each sample site, possibly as resting stages, is an important component in explaining the observed genetic structure.

Iron acquisition and allocation in stramenopile algae.

The essential element iron has a low biological availability in the surface ocean where photosynthetic organisms live. Recent advances in our understanding of iron acquisition mechanisms in brown algae and diatoms (stramenopile algae) show the importance of the reduction of ferric to ferrous iron prior to, or during, transport in the uptake process. The uses of iron in photosynthetic stramenopiles resembles that in other oxygenic organisms, although (with the exception of the diatom Thalassiosira oceanica from an iron-deficient part of the ocean) they lack plastocyanin, instead using cytochrome c 6, This same diatom further economizes genotypically on the use of iron in photosynthesis by decreasing the expression of photosystem I, cytochrome c 6, and the cytochrome b 6 f complex per cell and per photosystem II relative to the coastal Thalassiosira pseudonana; similar changes occur phenotypically in response to iron deficiency in other diatoms such as Phaeodactylum tricornutum. In some diatoms grown under iron-limiting conditions, essentially all of the iron in the cells can be accounted for by the iron occurring in catalytic proteins. However, stramenopiles can store iron. Genomic studies show that pennate, but not centric, diatoms have the iron storage protein ferritin. While Mössbauer and X-ray analysis of (57)Fe-labelled Ectocarpus siliculosus shows iron in an amorphous mineral phase resembling the core of ferritin, the genome shows no protein with significant sequence similarity to ferritin.

Potential of photoautotrophic microbial organisms in deciphering forensic issues.

In forensic pathology, solving the crime mystery of death due to drowning still remains a challenging issue. The amalgamation of autopsy findings and comparative study of diatoms recovered from the victim's body organs and suspected drowning site help to decipher the cause of death due to drowning or post-mortem immersion. Since the correct interpretation of the cause of death is an important criterion to provide justice to the victim, therefore, the main objective of our study is to throw light on the application of photoautotrophic micro-algal organisms, known as Diatoms, in solving seven cases of victims whose bodies were recovered from various water bodies of Himachal Pradesh, India. The diatom test was conducted by using reverse aqua regia solution (15 ml HNO3: 5 ml HCl) on the bone marrow extracted from the organs and water samples respectively. The informative outcomes of the experimental analysis demonstrated that the diatom test acts as a beneficial adjunct to solve drowning-related crimes where the exact cause of death remains hidden even after performing an autopsy of the victims. The protocol followed by the authors can be used conveniently to recover diatoms from bone marrow as well as from water samples. Our results showed that the maximum cases were of death due to accidental drowning but for one case of suicidal drowning in extremely cold water.

New and Interesting Taxa from the Diatom Genus Gomphonema Ehrenberg in Shallow, Nearshore Sites on the Eastern Coast of Lake Baikal.

During this investigation, sixteen species from the genus Gomphonema were found in a shallow bay (near Oimur, Kabansky District) located on the eastern shore of Lake Baikal. Eleven of these taxa have been described previously. Five species are described as new to science. We provide ecological information about these Gomphonema species, including their distribution within and outside Lake Baikal, and compare and contrast the new taxa with morphologically similar congeners. The diversity of morphologies present in Lake Baikal among the Gomphonema species suggests there has been both in-lake speciation as well as the introduction of various groups of species. The idea that Lake Baikal may support a high number of endemics by their partitioning niches based on depth is supported by the different groups of gomphonemoid diatoms present.

The Genus Pinnularia Ehrenberg (Bacillariophyta) from the Transbaikal Area (Russia, Siberia): Description of Seven New Species on the Basis of Morphology and Molecular Data with Discussion of the Phylogenetic Position of Caloneis.

Seven Pinnularia species from the Transbaikal area, Russia, are described as new for science. These are P. baicalgenkalii, P. baicalflexuosa, P. microfrauenbergiana, P. pergrunowii, P. siberiosinistra, P. baicalodivergens, and P. baicalislandica. All species are described by original LM and SEM microphotographs and molecular phylogeny. We provide comparisons between the taxa and document variability in the features found in the species. The number of Pinnularia species in the Transbaikal area is the largest number of species of the genus anywhere in the world.

A review of the taxonomic diversity, host-parasite interactions, and experimental research on chytrids that parasitize diatoms.

Diatoms (Bacillariophyta) are a major source of primary production on Earth, generating between 1/4 to 1/2 of all oxygen. They are found in almost all bodies of water, the ice of mountains, the arctic and the antarctic, and soils. Diatoms are also a major source of food in aquatic systems, a key component of the silica cycle, and are carbon capturers in oceans. Recently, diatoms have been examined as sources of biofuels, food, and other economic boons. Chytrids are members of the Kingdom fungi comprising, at a minimum, Chytridiomycota, Blastocladiomycota, and Neocallimastigales. Most chytrids are saprobes, plant pathogens, or parasites, and play an important role in aquatic ecosystems. Chytrid parasitism of diatoms has been reported to cause epidemics of over 90% fatality, though most of the information regarding these epidemics is limited to interactions between just a few hosts and parasites. Given the ubiquity of diatoms, their importance in natural and economic systems, and the massive impact epidemics can have on populations, the relative lack of knowledge regarding parasitism by chytrids is alarming. Here we present a list of the firsthand accounts of diatoms reported parasitized by chytrids. The list includes 162 named parasitic chytrid-diatom interactions, with 63 unique chytrid taxa from 11 genera, and 74 unique diatom taxa from 28 genera. Prior to this review, no list of all documented diatom-chytrid interactions existed. We also synthesize the currently known methods of infection, defense, and experiments examining diatoms and chytrids, and we document the great need for work examining both a greater breadth of taxonomic diversity of parasites and hosts, and a greater depth of experiments probing their interactions. This resource is intended to serve as a building block for future researchers studying diatom-parasite interactions and global planktonic communities in both fresh and marine systems.

Study of terrestrial diatoms in corticolous assemblages from deciduous trees in Central Europe with descriptions of two new Luticola D.G.Mann taxa.

Although many studies have examined the algae associated with various habitats in tree trunks, the diatoms in these environments are still poorly studied. Studies of corticolous algae mainly focus on green algae and cyanobacteria, which are usually immediately visible, while diatoms are mostly overlooked or not reported. During the research, 143 species of diatoms were identified, including two new representatives of the genus Luticola: L.bryophilasp. nov. with relatively large central area and short distal raphe endings and L.confusasp. nov. characterized by the presence of small depressions on central raphe endings. Both are described herein based on light and scanning electron microscopy observations and compared to similar taxa based on literature data. Basic morphological data for almost all the diatom taxa are noted, and their habitat requirements, and photographic documentation are also presented. The present research showed that the occurrence of diatom assemblages on tree trunks is influenced by various factors like host tree species, the area where the host tree grows, and the availability of suitable microhabitats within the trunk. However, the species composition of this assemblages depends mainly on the tree species.

Enhanced fixation of dissolved inorganic carbon by algal-bacterial aerobic granular sludge during treatment of low-organic-content wastewater.

The microalgae-based wastewater treatment technologies are believed to contribute to carbon neutrality. This study investigated the inorganic carbon fixation performance in the algal-bacterial aerobic granular sludge (A-BAGS) process under cultivation at different concentrations of organic carbon (OC) and inorganic carbon (IC). The results indicated that A-BAGS in treating wastewater containing organics of 77 mg-C/L contributed little to the fixation of inorganic carbon, while the highest inorganic carbon removal efficiency of 50 % was achieved at the influent IC of 100 mg/L and OC of 7 mg/L. This high IC condition contributed to enhanced biomass growth rate and enhanced extracellular polymeric substances, while it did not affect the granular stability and nitrification efficiency. The microbial diversity was also largely enhanced. The results demonstrated the great potential of A-BAGS for simultaneous resource recovery in wastewater and inorganic carbon fixation, while operation conditions need to be further optimized.

Taxonomy and Distribution of the Genus Sellaphora Mereschowsky (Bacillariophyceae: Sellaphoraceae) in Aquatic Ecosystems of Mongolia.

During our investigation, 28 species belonging to the genus Sellaphora Mereschowsky were found in different types of water bodies and streams in Mongolia. Of these, 14 species have already been described and/or shown earlier in the flora of Mongolia, and we found 14 species for the first time. One new taxonomic combination has been proposed: Sellaphora glomus (J.R. Carter and A.E. Bailey-Watts) Glushchenko and Kulikovskiy comb. nov. One species is proposed for description as new to science: Sellaphora dorofeyukae Glushchenko and Kulikovskiy sp. nov. All species were illustrated with original light micrographs, and a number of species were also illustrated with scanning electronic photographs. The distribution of the identified species within Mongolia and the world is discussed.

A Voucher Flora of Diatoms from Fens in the Tanana River Floodplain, Alaska.

Climate change and human activities may alter the structure and function of boreal peatlands by warming waters and changing their hydrology. Diatoms can be used to assess or track these changes. However, effective biomonitoring requires consistent, reliable identification. To address this need, this study developed a diatom voucher flora of species found across a boreal fen gradient (e.g., vegetation) in interior Alaskan peatlands. Composite diatom samples were collected bi-weekly from three peatland complexes over the 2017 summer. The morphological range of each taxon was imaged. The fens contained 184 taxa across 38 genera. Eunotia (45), Gomphonema (23), and Pinnularia (20) commonly occurred in each peatland. Tabellaria was common in the rich and moderate fen but sparse in the poor fen. Eunotia showed the opposite trend. Approximately 11% of species are potentially novel and 25% percent matched those at risk or declining in status on the diatom Red List (developed in Germany), highlighting the conservation value of boreal wetlands. This voucher flora expands knowledge of regional diatom biodiversity and provides updated, verifiable taxonomic information for inland Alaskan diatoms, building on Foged's 1981 treatment. This flora strengthens the potential to effectively track changes in boreal waterways sensitive to climate change and anthropogenic stressors.

Comparative Analysis of Bacillariophyceae Chloroplast Genomes Uncovers Extensive Genome Rearrangements Associated with Speciation.

The Bacillariophyceae is a species-rich, ecologically significant class of Bacillariophyta. Despite their critical importance in marine ecosystems as primary producers and in the development of harmful algal blooms (HABs), taxonomic research on Bacillariophyceae species has been hindered because of their limited morphological features, plasticity of morphologies, and the low resolution of common molecular markers. Hence molecular markers with improved resolution are urgently needed. Organelle genomes, which can be constructed efficiently with the recent development of high throughput DNA sequencing technologies and the advancement of bioinformatics tools, have been proposed as super barcodes for their higher resolution for distinguishing different species and intra-species genomic variations. In this study, we tested the value of full-length chloroplast genomes (cpDNAs) as super barcodes for distinguishing diatom species, by constructing cpDNAs of 11 strains of the class Bacillariophyceae, including Nitzschia ovalis, Nitzschia traheaformis, Cylindrotheca spp., Psammodictyon constrictum, Bacillaria paxillifer, two strains of Haslea tsukamotoi, Haslea avium, Navicula arenaria, and Pleurosigma sp. Comparative analysis of cpDNAs revealed that cpDNAs were not only adequate for resolving different species, but also for enabling recognition of high levels of genome rearrangements between cpDNAs of different species, especially for species of the genera Nitzschia, Cylindrotheca, Navicula and Haslea. Additionally, comparative analysis suggested that the positioning of species in the genus Haslea should be transferred to the genus Navicula. Chloroplast genome-based evolutionary analysis suggested that the Bacillariophyceae species first appeared during the Cretaceous period and the diversity of species rose after the mass extinction about 65 Mya. This study highlighted the value of cpDNAs in research on the biodiversity and evolution of Bacillariophyceae species, and, with the construction of more cpDNAs representing additional genera, deeper insight into the biodiversity and evolutionary relationships of Bacillariophyceae species will be gained.

Molecular and Morphological Investigations of Two Giant Diatom Cymbella Species from the Transbaikal Area (Russia, Siberia) with Comments on Their Distributions.

For the first time, a giant diatom species of the genus Cymbella from Lake Baikal was studied using molecular methods. Molecular and morphological investigations allowed to us to described one new species, Cymbella baicalaspera Glushchenko, Kulikovskiy and Kociolek sp. nov. This species is both morphologically similar and phylogenetically close to a second giant Cymbella species that we investigated here, identified by us as Cymbella himalaspera Jüttner and Van de Vijver in Jüttner et al. 2010. This species was first described from Nepal on the basis of a morphological investigation. Small morphological differences exist between the type population and specimens from Lake Baikal, but otherwise the two are identical. These very interesting results show that some Baikalian diatoms can be distributed more widely and are not only endemic to this ancient lake. Similarity between Cymbella baicalaspera sp. nov. and Cymbella himalaspera on the basis of both morphological features and their close phylogenetic relationships suggested by molecular data indicate they are sister species and an example of sympatric speciation. These results also suggest an early development of a species flock. This species group warrants additional research in terms of. their diversification and biogeography.

Draft genome assembly and sequencing dataset of the marine diatom Skeletonema cf. costatum RCC75.

Diatoms (Bacillariophyceae) are a major constituent of the phytoplankton and have a universally recognized ecological importance. Between 1,000 and 1,300 diatom genera have been described in the literature, but only 10 nuclear genomes have been published and made available to the public up to date. Skeletonema costatum is a cosmopolitan marine diatom, principally occurring in coastal regions, and is one of the most abundant members of the Skeletonema genus. Here we present a draft assembly of the Skeletonema cf. costatum RCC75 genome, obtained from PacBio and Illumina NovaSeq data. This dataset will expand the knowledge of the Bacillariophyceae genetics and contribute to the global understanding of phytoplankton's physiological, ecological, and environmental functioning.

Characterization of a CO2-Concentrating Mechanism with Low Sodium Dependency in the Centric Diatom Chaetoceros gracilis.

Microalgae induce a CO2-concentrating mechanism (CCM) to overcome CO2-limiting stress in aquatic environments by coordinating inorganic carbon (Ci) transporters and carbonic anhydrases (CAs). Two mechanisms have been suggested to facilitate Ci uptake from aqueous media: Na+-dependent HCO3- uptake by solute carrier (SLC) family transporters and accelerated dehydration of HCO3- to CO2 by external CA in model diatoms. However, studies on ecologically and industrially important diatoms including Chaetoceros gracilis, a common food source in aquacultures, are still limited. Here, we characterized the CCM of C. gracilis using inhibitors and growth dependency on Na+ and CO2. Addition of a membrane-impermeable SLC inhibitor, 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS), or the transient removal of Na+ from the culture medium did not impair photosynthetic affinity for Ci in CO2-limiting stress conditions, but addition of a membrane-impermeable CA inhibitor, acetazolamide, decreased Ci affinity to one-third of control cultures. In culture medium containing 0.23 mM Na+ C. gracilis grew photoautotrophically by aeration with air containing 5% CO2, but not with the air containing 0.04% CO2. These results suggested that C. gracilis utilizes external CAs in its CCM to elevate photosynthetic affinity for Ci rather than plasma-membrane SLC family transporters. In addition, it is possible that low level of Na+ may support the CCM in processes other than Ci-uptake at the plasma membrane specifically in CO2-limiting conditions. Our findings provide insights into the diversity of CCMs among diatoms as well as basic information to optimize culture conditions for industrial applications.

Achnanthidium tinea sp. nov. - a new monoraphid diatom (Bacillariophyceae) species, described on the basis of molecular and morphological approaches.

A new monoraphid diatom species Achnanthidium tinea Tseplik, Kulikovskiy, Kociolek & Maltsev, sp. nov. is described from Indonesia. The species is described on the basis of molecular and morphological analyses. According to molecular data the new species belongs to the clade that includes strains of Achnanthidium minutissimum, Achnanthidium saprophilum and Achnanthidium digitatum. Morphologically, the new species differs quite significantly from other species of the same genus because of linear-elliptic valves with almost parallel sides and strongly radiate striae and a butterfly-shaped fascia on the raphe valve. The morphology and phylogeny of the new species are discussed, and thoughts on the current state of the taxonomy of the genus Achnanthidium are expressed. Our work shows the importance of using molecular data in diatom systematics and also demonstrates the need to investigate rarely studied regions of our planet.