Endolithic Fungal Diversity in Antarctic Oligocene Rock Samples Explored Using DNA Metabarcoding.

In this study, we evaluated the fungal diversity present associated with cores of Oligocene rocks using a DNA metabarcoding approach. We detected 940,969 DNA reads grouped into 198 amplicon sequence variants (ASVs) representing the phyla Ascomycota, Basidiomycota, Mortierellomycota, Chytridiomycota, Mucoromycota, Rozellomycota, Blastocladiomycota, Monoblepharomycota, Zoopagomycota, Aphelidiomycota (Fungi) and the fungal-like Oomycota (Stramenopila), in rank abundance order. Pseudogymnoascus pannorum, Penicillium sp., Aspergillus sp., Cladosporium sp., Aspergillaceae sp. and Diaporthaceae sp. were assessed to be dominant taxa, with 22 fungal ASVs displaying intermediate abundance and 170 being minor components of the assigned fungal diversity. The data obtained displayed high diversity indices, while rarefaction indicated that the majority of the diversity was detected. However, the diversity indices varied between the cores analysed. The endolithic fungal community detected using a metabarcoding approach in the Oligocene rock samples examined contains a rich and complex mycobiome comprising taxa with different lifestyles, comparable with the diversity reported in recent studies of a range of Antarctic habitats. Due to the high fungal diversity detected, our results suggest the necessity of further research to develop strategies to isolate these fungi in culture for evolutionary, physiological, and biogeochemical studies, and to assess their potential role in biotechnological applications.

Fungal diversity present in snow sampled in summer in the north-west Antarctic Peninsula and the South Shetland Islands, Maritime Antarctica, assessed using metabarcoding.

We assessed the fungal diversity present in snow sampled during summer in the north-west Antarctic Peninsula and the South Shetland Islands, maritime Antarctica using a metabarcoding approach. A total of 586,693 fungal DNA reads were obtained and assigned to 203 amplicon sequence variants (ASVs). The dominant phylum was Ascomycota, followed by Basidiomycota, Mortierellomycota, Chytridiomycota and Mucoromycota. Penicillium sp., Pseudogymnoascus pannorum, Coniochaeta sp., Aspergillus sp., Antarctomyces sp., Phenoliferia sp., Cryolevonia sp., Camptobasidiaceae sp., Rhodotorula mucilaginosa and Bannozyma yamatoana were assessed as abundant taxa. The snow fungal diversity indices were high but varied across the different locations sampled. Of the fungal ASVs detected, only 28 were present all sampling locations. The 116 fungal genera detected in the snow were dominated by saprotrophic taxa, followed by symbiotrophic and pathotrophic. Our data indicate that, despite the low temperature and oligotrophic conditions, snow can host a richer mycobiome than previously reported through traditional culturing studies. The snow mycobiome includes a complex diversity dominated by cosmopolitan, cold-adapted, psychrophilic and endemic taxa. While saprophytes dominate this community, a range of other functional groups are present.

Fungal and fungal-like diversity present in ornithogenically influenced maritime Antarctic soils assessed using metabarcoding.

We assessed soil fungal and fungal-like diversity using metabarcoding in ornithogenically influenced soils around nests of the bird species Phalacrocorax atriceps, Macronectes giganteus, Pygoscelis antarcticus, and Pygoscelis adelie on the South Shetland Islands, maritime Antarctic. A total of 1,392,784 fungal DNA reads was obtained and assigned to 186 amplicon sequence variants (ASVs). The dominant fungal phylum was Ascomycota, followed by Basidiomycota, Chytridiomycota, Blastocladiomycota, Rozellomycota, Mortierellomycota, Monoblepharomycota, Aphelidiomycota, Basidiobolomycota, Mucoromycota, and the fungal-like Oomycota (Stramenopila), in rank order. Antarctomyces sp., Blastocladiomycota sp., Pseudogymnoascus pannorum, Microascaceae sp., Mortierella sp., Lobulomycetales sp., Sordariomycetes sp., Fungal sp., Rhizophydiales sp., Pseudeurotiaceae sp., Chytridiomycota sp. 1, Filobasidiella sp., Tausonia pullulans, Betamyces sp., and Leucosporidium sp. were the most abundant assigned taxa. The fungal assemblages present in the different ornithogenically influenced soils displayed different diversity indices. However, in general, we detected high fungal diversity and few taxa shared between the samples. Despite the polyextreme environmental conditions experienced in these Antarctic soils, the metabarcoding approach detected a rich and complex fungal community dominated by saprophytes, but with some pathogenic taxa also present. The community was dominated by psychrophilic and psychrotolerant taxa, some apparently endemic to Antarctica, and those identified only at higher taxonomic levels, which may represent currently undescribed fungi. The mycobiome detected included taxa characterized by different ecological roles, including saprotrophic, human- and animal-associated, phytopathogenic, mutualistic, and cosmopolitan. These fungi may potentially be dispersed by birds or in the air column over great distances, including between different regions within Antarctica and from South America, Africa, and Oceania.

An outsider on the Antarctic Peninsula: A new record of the non-native moth Plodia interpunctella (Lepidoptera: Pyralidae).

We report the first record of the microlepidopteran Plodia interpunctella beyond the South Shetland Islands at the Chilean Yelcho scientific station (64°52'33.1428″ S; 63°35'1.9572″ W), Doumer Island, close to the west coast of the Antarctic Peninsula. It is notable that P. interpunctella, a globally distributed stored product pest species, exhibits a remarkable capacity for prolonged viability within food storage facilities. The dual challenges of food transportation and storage in the context of Antarctica's challenging operational conditions may have facilitated P. interpunctella's initial arrival to the Antarctic region. Non-perishable food items, such as grains, flour and rice, provide practical options for the bulk food transportation and storage required in the long-term operation of Antarctic research stations. The presence of P. interpunctella in Antarctica, even if restricted to synanthropic environments within buildings, is a clear threat to Antarctic biodiversity, not only through being an invasive species itself but also as a potential vector for other non-native species (bacteria, acari, between others.), which could carry diseases to the native species.

Correction: A stomata classification and detection system in microscope images of maize cultivars.

[This corrects the article DOI: 10.1371/journal.pone.0258679.].

Performance of ChatGPT on the Brazilian Radiology and Diagnostic Imaging and Mammography Board Examinations.

This prospective exploratory study conducted from January 2023 through May 2023 evaluated the ability of ChatGPT to answer questions from Brazilian radiology board examinations, exploring how different prompt strategies can influence performance using GPT-3.5 and GPT-4. Three multiple-choice board examinations that did not include image-based questions were evaluated: (a) radiology and diagnostic imaging, (b) mammography, and (c) neuroradiology. Five different styles of zero-shot prompting were tested: (a) raw question, (b) brief instruction, (c) long instruction, (d) chain-of-thought, and (e) question-specific automatic prompt generation (QAPG). The QAPG and brief instruction prompt strategies performed best for all examinations (P < .05), obtaining passing scores (≥60%) on the radiology and diagnostic imaging examination when testing both versions of ChatGPT. The QAPG style achieved a score of 60% for the mammography examination using GPT-3.5 and 76% using GPT-4. GPT-4 achieved a score up to 65% in the neuroradiology examination. The long instruction style consistently underperformed, implying that excessive detail might harm performance. GPT-4's scores were less sensitive to prompt style changes. The QAPG prompt style showed a high volume of the "A" option but no statistical difference, suggesting bias was found. GPT-4 passed all three radiology board examinations, and GPT-3.5 passed two of three examinations when using an optimal prompt style. Keywords: ChatGPT, Artificial Intelligence, Board Examinations, Radiology and Diagnostic Imaging, Mammography, Neuroradiology © RSNA, 2023 See also the commentary by Trivedi and Gichoya in this issue.

Investigating non-fungal eukaryotic diversity in snow in the Antarctic Peninsula region using DNA metabarcoding.

Snow is a unique microhabitat, despite being a harsh environment, multiple life forms have adapted to survive in it. While algae, bacteria and fungi are dominant microorganisms in Antarctic snow, little is known about other organisms that may be present in this habitat. We used metabarcoding to investigate DNA sequence diversity of non-fungal eukaryotes present in snow obtained from six different sites across the Maritime Antarctica. A total of 20 taxa were assigned to obtained sequences, representing five Kingdoms (Chromista, Protozoa, Viridiplantae and Metazoa) and four phyla (Ciliophora, Cercozoa, Chlorophyta and Cnidaria). The highest diversity indices were detected in Trinity Peninsula followed by Robert Island, Arctowski Peninsula, Deception Island, King George Island and Snow Island. The most abundant assignments were to Trebouxiophyceae, followed by Chlamydomonas nivalis and Chlamidomonadales. No taxa were detected at all sites. Three potentially new records for Antarctica were detected: two Ciliophora (Aspidisca magna and Stokesia sp.) and the green algae Trebouxia potteri. Our data suggested that similarities found between the sites may be more related with snow physicochemical properties rather than geographic proximity or latitude. This study provides new insights into the diversity and distribution of eukaryotic organisms in Antarctic snow.

Integrating public engagement to intensify pollination services through ecological restoration.

Globally, human activities impose threats to nature and the provision of ecosystem services, such as pollination. In this context, ecological restoration provides opportunities to create managed landscapes that maximize biodiversity conservation and sustainable agriculture, e.g., via provision of pollination services. Managing pollination services and restoration opportunities requires the engagement of distinct stakeholders embedded in diverse social institutions. Nevertheless, frameworks toward sustainable agriculture often overlook how stakeholders interact and access power in social arenas. We present a perspective integrating pollination services, ecological restoration, and public engagement for biodiversity conservation and agricultural production. We highlight the importance of a comprehensive assessment of pollination services, restoration opportunities identification, and a public engagement strategy anchored in institutional analysis of the social arenas involved in restoration efforts. Our perspective can therefore guide the implementation of practices from local to country scales to enhance biodiversity conservation and sustainable agriculture.

Artificial Intelligence in Radiology: A Private Practice Perspective From a Large Health System in Latin America.

In the field of radiology, the use of artificial intelligence (AI) is increasing. Even though healthcare facilities are interested in using this technology, having success with an AI project can be challenging. There is a myriad of AI solutions today, and comparing them can be challenging. Moreover, the implementation process involves alignment with many different areas. In our institution, we have been testing, developing, deploying, and monitoring AI solutions for the last four years. This article intends to share our experience and highlight the most important points to ensure a successful project based on our experience in the setting of a large private practice in Latin America.

Soil Fungal Diversity and Ecology Assessed Using DNA Metabarcoding along a Deglaciated Chronosequence at Clearwater Mesa, James Ross Island, Antarctic Peninsula.

We studied the fungal diversity present in soils sampled along a deglaciated chronosequence from para- to periglacial conditions on James Ross Island, north-east Antarctic Peninsula, using DNA metabarcoding. A total of 88 amplicon sequence variants (ASVs) were detected, dominated by the phyla Ascomycota, Basidiomycota and Mortierellomycota. The uncommon phyla Chytridiomycota, Rozellomycota, Monoblepharomycota, Zoopagomycota and Basidiobolomycota were detected. Unknown fungi identified at higher hierarchical taxonomic levels (Fungal sp. 1, Fungal sp. 2, Spizellomycetales sp. and Rozellomycotina sp.) and taxa identified at generic and specific levels (Mortierella sp., Pseudogymnoascus sp., Mortierella alpina, M. turficola, Neoascochyta paspali, Penicillium sp. and Betamyces sp.) dominated the assemblages. In general, the assemblages displayed high diversity and richness, and moderate dominance. Only 12 of the fungal ASVs were detected in all chronosequence soils sampled. Sequences representing saprophytic, pathogenic and symbiotic fungi were detected. Based on the sequence diversity obtained, Clearwater Mesa soils contain a complex fungal community, including the presence of fungal groups generally considered rare in Antarctica, with dominant taxa recognized as cold-adapted cosmopolitan, endemic, saprotrophic and phytopathogenic fungi. Clearwater Mesa ecosystems are impacted by the effects of regional climatic changes, and may provide a natural observatory to understand climate change effects over time.

DNA Metabarcoding Reveals Cryptic Diversity in Forest Soils on the Isolated Brazilian Trindade Island, South Atlantic.

Located 1140 km from the South American coastline in the South Atlantic Ocean and with an age of 4 million years, Trindade Island is the most recent volcanic component of Brazilian territory. Its original native vegetation has been severely damaged by human influence, in particular through the introduction of exotic grazing animals such as goats. However, since the complete eradication of goats and other feral animals in the late 1990s, the island's vegetation has been recovering, and even some endemic species that had been considered extinct have been rediscovered. In this study, we set out to characterize the contemporary cryptic diversity in soils of the recovering native forest of Trindade Island using metabarcoding by high throughput sequencing (HTS). The sequence diversity obtained was dominated by microorganisms, including three domains (Bacteria, Archaea, and Eukarya) and five kingdoms (Fungi, Metazoa, Protozoa, Chromista, and Viridiplantae). Bacteria were represented by 20 phyla and 116 taxa, with Archaea by only one taxon. Fungi were represented by seven phyla and 250 taxa, Viridiplantae by five phyla and six taxa, Protozoa by five phyla and six taxa, Metazoa by three phyla and four taxa and Chromista by two phyla and two taxa. Even after the considerable anthropogenic impacts and devastation of the island's natural forest, our sequence data reveal the presence of a rich and complex diversity of microorganisms, invertebrates, and plants and provide important baseline biodiversity information that will contribute to ecological restoration efforts on the island.

Fungal and fungal-like diversity in marine sediments from the maritime Antarctic assessed using DNA metabarcoding.

We assessed the fungal and fungal-like sequence diversity present in marine sediments obtained in the vicinity of the South Shetland Islands (Southern Ocean) using DNA metabarcoding through high-throughput sequencing (HTS). A total of 193,436 DNA reads were detected in sediment obtained from three locations: Walker Bay (Livingston Island) at 52 m depth (48,112 reads), Whalers Bay (Deception Island) at 151 m (104,704) and English Strait at 404 m (40,620). The DNA sequence reads were assigned to 133 distinct fungal amplicon sequence variants (ASVs) representing the phyla Ascomycota, Basidiomycota, Mortierellomycota, Chytridiomycota, Glomeromycota, Monoblepharomycota, Mucoromycota and Rozellomycota and the fungal-like Straminopila. Thelebolus balaustiformis, Pseudogymnoascus sp., Fungi sp. 1, Ciliophora sp., Agaricomycetes sp. and Chaetoceros sp. were the dominant assigned taxa. Thirty-eight fungal ASVs could only be assigned to higher taxonomic levels, and may represent taxa not currently included in the available databases or represent new taxa and/or new records for Antarctica. The total fungal community displayed high indices of diversity, richness and moderate to low dominance. However, diversity and taxa distribution varied across the three sampling sites. In Walker Bay, unidentified fungi were dominant in the sequence assemblage. Whalers Bay sediment was dominated by Antarctic endemic and cold-adapted taxa. Sediment from English Strait was dominated by Ciliophora sp. and Chaetoceros sp. These fungal assemblages were dominated by saprotrophic, plant and animal pathogenic and symbiotic taxa. The detection of an apparently rich and diverse fungal community in these marine sediments reinforces the need for further studies to characterize their richness, functional ecology and potential biotechnological applications.

Antarctic organisms as a source of antimicrobial compounds: a patent review.

Currently, antimicrobial resistance has become a global public health problem, which has made the need for new antimicrobial compounds to deal with resistant infections an emergency. However, environments that once offered so many innovative molecules, now already exhaustively exploited, do not meet this need. In this context, a geographically isolated, under-explored and extreme environment, such as Antarctica, which holds organisms with unique physiological and biochemical characteristics, assumes great importance as a potential source of new compounds with antimicrobial activity. In this patent review, we investigate the state of technological development in the field of antimicrobial compounds obtained from Antarctic organisms, highlighting the main countries and researchers active in the field, the species utilized, the compounds obtained, and their possible therapeutic applications. As results, few patent documents were found, however they encompass a wide diversity of compounds and species, indicating a great antimicrobial potential present in Antarctic biota, including compounds active against the most important human pathogenic microorganisms, such as including methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus spp. and multi-resistant Mycobacterium tuberculosis. Furthermore, due to the increasing trend in patent applications, a significant rise in the number of patents in this area is expected in the coming years.

Using metabarcoding to assess Viridiplantae sequence diversity present in Antarctic glacial ice.

Antarctica contains most of the glacial ice on the planet, a habitat that is largely unexplored by biologists. Recent warming in parts of Antarctica, particularly the Antarctic Peninsula region, is leading to widespread glacial retreat, releasing melt water and, potentially, contained biological material and propagules. In this study, we used a DNA metabarcoding approach to characterize Viridiplantae DNA present in Antarctic glacial ice. Ice samples from six glaciers in the South Shetland Islands and Antarctic Peninsula were analysed, detecting the presence of DNA representing a total of 16 taxa including 11 Chlorophyta (green algae) and five Magnoliophyta (flowering plants). The green algae may indicate the presence of a viable algal community in the ice or simply of preserved DNA, and the sequence diversity assigned included representatives of Chlorophyta not previously recorded in Antarctica. The presence of flowering plant DNA is most likely to be associated with pollen or tissue fragments introduced by humans.

Diversity of Fungi Present in Permafrost in the South Shetland Islands, Maritime Antarctic.

We assess the fungal diversity present in permafrost from different islands in the South Shetland Islands archipelago, maritime Antarctic, using next-generation sequencing (NGS). We detected 1,003,637 fungal DNA reads representing, in rank abundance order, the phyla Ascomycota, Mortierellomycota, Basidiomycota, Chytridiomycota, Rozellomycota, Mucoromycota, Calcarisporiellomycota and Zoopagomycota. Ten taxa were dominant these being, in order of abundance, Pseudogymnoascus appendiculatus, Penicillium sp., Pseudogymnoascus roseus, Penicillium herquei, Curvularia lunata, Leotiomycetes sp., Mortierella sp. 1, Mortierella fimbricystis, Fungal sp. 1 and Fungal sp. 2. A further 38 taxa had intermediate abundance and 345 were classified as rare. The total fungal community detected in the permafrost showed high indices of diversity, richness and dominance, although these varied between the sampling locations. The use of a metabarcoding approach revealed the presence of DNA of a complex fungal assemblage in the permafrost of the South Shetland Islands including taxa with a range of ecological functions among which were multiple animal, human and plant pathogenic fungi. Further studies are required to determine whether the taxa identified are present in the form of viable cells or propagules and which might be released from melting permafrost to other Antarctic habitats and potentially dispersed more widely.

Fungi in the Antarctic Cryosphere: Using DNA Metabarcoding to Reveal Fungal Diversity in Glacial Ice from the Antarctic Peninsula Region.

We assessed fungal diversity present in glacial from the Antarctic Peninsula using DNA metabarcoding through high-throughput sequencing (HTS). We detected a total of 353,879 fungal DNA reads, representing 94 genera and 184 taxa, in glacial ice fragments obtained from seven sites in the north-west Antarctic Peninsula and South Shetland Islands. The phylum Ascomycota dominated the sequence diversity, followed by Basidiomycota and Mortierellomycota. Penicillium sp., Cladosporium sp., Penicillium atrovenetum, Epicoccum nigrum, Pseudogymnoascus sp. 1, Pseudogymnoascus sp. 2, Phaeosphaeriaceae sp. and Xylaria grammica were the most dominant taxa, respectively. However, the majority of the fungal diversity comprised taxa of rare and intermediate relative abundance, predominately known mesophilic fungi. High indices of diversity and richness were calculated, along with moderate index of dominance, which varied among the different sampling sites. Only 26 (14%) of the total fungal taxa detected were present at all sampling sites. The identified diversity was dominated by saprophytic taxa, followed by known plant and animal pathogens and a low number of symbiotic fungi. Our data suggest that Antarctic glacial ice may represent a hotspot of previously unreported fungal diversity; however, further studies are required to integrate HTS and culture approaches to confirm viability of the taxa detected.

A stomata classification and detection system in microscope images of maize cultivars.

Plant stomata are essential structures (pores) that control the exchange of gases between plant leaves and the atmosphere, and also they influence plant adaptation to climate through photosynthesis and transpiration stream. Many works in literature aim for a better understanding of these structures and their role in the evolution process and the behavior of plants. Although stomata studies in dicots species have advanced considerably in the past years, even there is not much knowledge about the stomata of cereal grasses. Due to the high morphological variation of stomata traits intra- and inter-species, detecting and classifying stomata automatically becomes challenging. For this reason, in this work, we propose a new system for automatic stomata classification and detection in microscope images for maize cultivars based on transfer learning strategy of different deep convolution neural netwoks (DCNN). Our performed experiments show that our system achieves an approximated accuracy of 97.1% in identifying stomata regions using classifiers based on deep learning features, which figures out as a nearly perfect classification system. As the stomata are responsible for several plant functionalities, this work represents an important advance for maize research, providing an accurate system in replacing the current manual task of categorizing these pores on microscope images. Furthermore, this system can also be a reference for studies using images from different cereal grasses.

Fairy ring disease affects epiphytic algal assemblages associated with the moss Sanionia uncinata (Hedw.) Loeske (Bryophyta) on King George Island, Antarctica.

Since the nineteenth century, a ring-forming disease attacking Antarctic mosses has been reported. However, to date, only the effects on the mosses themselves are known. In this study, we used DNA metabarcoding to investigate the effects on the moss epiphytic algal community at different stages of disease progression. As the disease progressed, algal species richness decreased, although overall abundance was not significantly affected. Prasiolales appeared unaffected, whereas Ulotrichales were more sensitive. Trebouxiales dominated the advanced disease stage, suggesting a possible benefit from the disease, either through the elimination of competition or creation of new niches. Infection is responsible for moss death, leading to habitat loss for other organisms, but pathogenic effects on algae cannot be ruled out. Our data indicate that the disease not only impacts mosses but also other groups, potentially resulting in loss of Antarctic biodiversity. This study provides the first report of the disease effects on epiphytic algal communities of Antarctic bryophytes.

Ecological succession of fungal and bacterial communities in Antarctic mosses affected by a fairy ring disease.

We evaluated fungal and bacterial diversity in an established moss carpet on King George Island, Antarctica, affected by 'fairy ring' disease using metabarcoding. A total of 127 fungal and 706 bacterial taxa were assigned. Ascomycota dominated the fungal assemblages, followed by Basidiomycota, Rozellomycota, Chytridiomycota, Mortierellomycota and Monoblepharomycota. The fungal community displayed high indices of diversity, richness and dominance, which increased from healthy through infected to dead moss samples. A range of fungal taxa were more abundant in dead rather than healthy or fairy ring moss samples. Bacterial diversity and richness were greatest in healthy moss and least within the infected fairy ring. The dominant prokaryotic phyla were Actinobacteriota, Proteobacteria, Bacteroidota and Cyanobacteria. Cyanophyceae sp., whilst consistently dominant, were less abundant in fairy ring samples. Our data confirmed the presence and abundance of a range of plant pathogenic fungi, supporting the hypothesis that the disease is linked with multiple fungal taxa. Further studies are required to characterise the interactions between plant pathogenic fungi and their host Antarctic mosses. Monitoring the dynamics of mutualist, phytopathogenic and decomposer microorganisms associated with moss carpets may provide bioindicators of moss health.