Antarctica has one of the most sensitive ecosystems to the negative effects of Persistent Organic Pollutants (POPs) on its biodiversity. This is because of the lower temperatures and the persistence of POPs that promote their accumulation or even biomagnification. However, the impact of POPs on vascular plants is unknown. Moreover, fungal symbionts could modulate the effects on host plants to cope with this stress factor. This study investigates the molecular and ecophysiological responses of the Sub-Antarctic and Antarctic plant Colobanthus quitensis to POPs in different populations along a latitudinal gradient (53 degrees- 67 degrees S), emphasizing the role of endophytic fungi. The results show that exposure of POPs in C. quitensis generates oxidative stress and alters its ecophysiological performance. Nevertheless, C. quitensis in association with fungal endophytes and POPs exposure, shows lower lipid peroxidation, higher proline content and higher photosynthetic capacity, as well as higher biomass and survival percentage, compared to plants in the absence of fungal endophytes. On the other hand, the antarctic plant population (67 degrees S) with endophytic fungi presents better stress modulating upon POPs exposure. Endophytic fungi would be more necessary for plant performance towards higher latitudes with extreme conditions, contributing significantly to their general functional adaptation. We develop a transcriptomics analyses n the C. quitensis-fungal endophytes association from the Peninsula population. We observed that fungal endophytes promote tolerance to POPs stress through upregulated genes for the redox regulation based on ascorbate and scavenging mechanisms (peroxidases, MDAR, VTC4, CCS), transformation (monooxygenases) and conjugation of compounds or metabolites (glutathione transferases, glycosyltransferases, S-transferases), and the storage or elimination of conjugates (ABC transporters, C and G family) that contribute to detoxification cell. This work highlights the contribution of endophytic fungi to plant resistance in situations of environmental stress, especially in extreme conditions such as in antarctica exposed to anthropogenic impact. The implications of these findings are relevant for the biosecurity of one of the last pristine bastions worldwide.

Northwestern Patagonia is inhabited by coastal dolphin species, including the Chilean (Cephalorhynchus eutropia) and Peale's dolphin (Lagenorhynchus australis), which use habitats under contrasting anthropogenic pressures. To explore whether the skin bacteriome of Chilean and Peale's dolphins reflects anthropogenic changes in their environment, we collected 122 skin biopsies, sequenced the V4 hypervariable region of the 16S rRNA gene, and investigated differences in the diversity, composition, and divergence of their bacterial communities across six different locations. Chilean and Peale's dolphins from the highly anthropized locations showed a remarkably different composition of the skin bacteriome compared to dolphins from other locations, with a high relative abundance of Vibrio and Halomonas. The bacteriomes of dolphins from highly anthropized locations were highly similar, with low uniqueness. Such homogenization of skin bacteriomes between dolphins may be an early warning sign of deviations from optimal health. The skin bacteriome of Chilean and Peale's dolphins proved to be a good indicator of the level of anthropization of their habitat.

Wastewater is a hotspot for viral diversity, harboring various microbial, plant, and animal viruses, including those that infect humans. However, the dynamics, resilience, and ecological roles of viral communities during treatment are largely unknown. In this study, we explored RNA virus ecogenomics using metagenomics from influent and effluent samples across three wastewater catchment areas in Chile, with a population of 7.05 million equivalent inhabitants. We identified 14,212 RNA-dependent RNA polymerase (RdRP)-coding sequences from the Orthornavirae kingdom, clustering into 4,989 viral species. Using extensive databases of 14,150 family-level representative sequences, we classified 90% of our sequences at the family level. Our analysis revealed that treatment reduced viral richness and evenness (Shannon index), but phylogenetic diversity remained unchanged. Effluents showed lower richness and evenness than influents with similar phylogenetic diversity. Species turnover, influenced by catchment area and treatment, accounted for 54% of sample dissimilarities (Weighted Unifrac). Biomarker analysis indicated that families like Astroviridae and Fiersviridae were more abundant in influents, while Reoviridae and Virgaviridae dominated effluents. This suggests that viral resistance to treatment varies and cannot be solely attributed to genome type, size, or morphology. We traced viral genomes through time and space, identifying sequences like the Pepper Mild Mottle Virus (PMMoV) from the Virgaviridae family over large distances and periods, highlighting its wastewater marker potential. High concentrations of human pathogens, such as Rotavirus (Reoviridae) and Human Astrovirus (Astroviridae), were found in both influents and effluents, stressing the need for continuous monitoring, especially for treated wastewater reuse.

Rhizosphere microbes establish functional interactions with their hosts, impacting plant fitness. To further understand plant effects on microbial composition and functional diversity, we present 52 metagenomes and 1,484 metagenome-assembled genomes (MAGs) from soil and the rhizosphere of Colobanthus quitensis and Deschampsia antarctica.

Semi-volatile organic compounds (SVOCs) are widely distributed across the globe, including polar regions. This study investigates the distribution and bioconcentration of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs) in soils and Colobanthus quitensis, while also estimating potential emission sources. Results indicated high concentrations of PAHs in soils and plants from the Sub-Antarctic region, while OCPs and PCBs were more prevalent in the Antarctic region, with higher contaminant concentrations found in soils than in plant tissues. Hexachlorobenzene (HCB) and dichlorodiphenyldichloroethylene (p,p′-DDE) were significantly higher in the Antarctic region, suggesting historical dichlorodiphenyltrichloroethane (DDT) use, while PCB 153 and 180 were the most representative PCBs in the Antarctic region. Phenanthrene (Phe) was the dominant PAH in both regions. The bioconcentration factor analysis from soils (BCFSoils) revealed potential anthropogenic influences for certain contaminants, including γ-hexachlorocyclohexane (γ-HCH) and PCB 9 in the Sub-Antarctic region, and HCB, p,p′-DDE, PCB 9, and benzo-naphtho-thiophene in the Antarctic region. However, compounds with higher hydrophobicity showed lower Bioconcentration factor (BCFSoils) values, indicating a tendency to accumulate in soil rather than plant tissues. This was consistent with the inverse relationship found between BCFSoils and the octanol-water partition coefficient (Log KOW). Diagnostic ratios of PAHs revealed a predominantly pyrogenic source in the Sub-Antarctic region, while a mixture of sources was observed in the Antarctic region.

Allergic rhinitis and asthma are two prevailing chronic airway diseases and serious public health concerns. Previous research has already described the role of the airway bacteriome in these two diseases, but almost no study so far has explored the mycobiome and its possible association to airway inflammation. Here we sequenced the internal transcribed spacers (ITS) 1 and 2 to characterize the oral mycobiome of 349 Portuguese children and young adults with allergic rhinitis alone (AR) or with asthma (ARAS), asthmatics (AS) and healthy controls (HC). Our genomic analyses showed that the two most abundant fungal phyla (Ascomycota and Basidiomycota) and 3–5 of the 14 most abundant fungal genera (Cladosporium, Aspergillus, Aleurina, Candida and Rhodotorula) in the mouth differed significantly (P ≤ 0.04) between both rhinitic groups and HC. However, none of the same taxa varied significantly between the three respiratory disease groups (AR, ARAS and AS). The oral mycobiomes of respiratory ill patients showed the highest intra-group diversity (microbial richness and evenness), while HC showed the lowest, with all alpha-diversity indices varying significantly (P ≤ 0.0424) between them. Similarly, all disease groups showed significant differences (P ≤ 0.0052) in microbial structure (i.e., beta-diversity indices) when compared to HC samples. Thirty metabolic pathways (PICRUSt2) were differentially abundant (Wald's test) between AR or ARAS and HC patients, but only one of them (D-galactose degradation I) was over abundant (log2 Fold Change >0.75) in the ARAS group. Spiec-Easi fungal networks varied greatly among groups, which suggests chronic respiratory allergic diseases may alter fungal connectivity in the mouth. This study increases our comprehension of the role of the oral mycobiome in allergy-related conditions. It shows for the first time that the oral mycobiota changes during health and allergic rhinitis (with and without asthma comorbidity) and highlights specific taxa, metabolic pathways and fungal interactions that may relate to chronic airway disease.

The high-latitude regions of Antarctica remain among the most remote, extreme, and least explored areas on Earth. Still, microbial life has been reported in these environments, with limited information on their genetic properties and functional capabilities. Although diverse autochthonous multidrug-resistant bacteria were found in Antarctic Peninsula soils, posing whether these soils could act as a source of resistance determinants that could emerge among pathogens, we still lack information regarding the resistome of areas closer to the South Pole. Moreover, no previous studies have evaluated the pathogenic potential of microbes inhabiting Antarctic soils. In this work, we combined metagenomic and culture-dependent approaches to investigate the microbial diversity, resistome, virulome, and mobile genetic elements (MGEs) in soils from Union Glacier, a cold desert in West Antarctica. Despite the extreme conditions, several bacterial phyla were found, predominating Actinomycetota and Pseudomonadota, with limited archaeal and fungal taxa. Contrastive with Ecology Glacier soils from King George Island, the Union Glacier soil bacterial community is significantly less diverse, mainly attributed to scarce moisture. We recovered >80 species-level representative genomes (SRGs) of predominant bacteria and an ammonia-oxidating nitrogen- and carbon-fixing archaeon from a novel species of Nitrosocosmicus. Several resistance and virulence genes were found in Union Glacier soils, similar to those in other Antarctic cold desert areas but significantly distinct from those observed in maritime Antarctica and other non-cryosphere biomes. Furthermore, we characterized bacterial isolates resistant to up to 24 clinical antibiotics, mainly Pseudomonas, Arthrobacter, Plantibacter, and Flavobacterium. Moreover, some isolates produced putative virulence factors, including siderophores, pyocyanins, and exoenzymes with hemolytic, lecithinase, protease, and DNAse activity. This evidence uncovers a largely unexplored resistome and virulome hosted by deep Antarctica's soil microbial communities and the presence of bacteria with pathogenic potential, highlighting the relevance of One Health approaches for environmental surveillance in this continent.

While the evidence supports a role for the soil microbiome to modulate the environmental tolerance of various plant species, its role in the invasion success remains seldom assessed. Here we show results from two complementary experiments aimed at understanding the role of the soil microbiome on the performance of T. officinale (dandelion) plants. Since the relative importance of soil microbiome on plant fitness can differ between native versus introduced origins, we conducted a full cross-transplant experiment to compare the plant performance from different origins (native/introduced) in their original or exotic soils, along with manipulated soil microbiome. In addition, since the relevance of soil microbiome for plant fitness can depend on the level of environmental stress, we compared the plant performance under different soil microbiome treatments in an introduced latitudinal gradient. We found positive effects of soil microbiome on performance traits for T. officinale plants from most of the evaluated populations, being particularly relevant for plants in the introduced range and under stressful conditions.

Escherichia coli is the leading cause of urinary tract infections (UTIs) in children and adults. The gastrointestinal tract is the primary reservoir of uropathogenic E. coli, which can be acquired from a variety of environmental exposures, including retail meat. In the current study, we used a novel statistical-genomic approach to estimate the proportion of pediatric UTIs caused by foodborne zoonotic E. coli strains. E. coli urine isolates were collected from DC residents aged 2 months to 17 years from the Children's National Medical Center Laboratory, 2013-2014. During the same period, E. coli isolates were collected from retail poultry products purchased from 15 sites throughout DC. A total of 52 urine and 56 poultry isolates underwent whole-genome sequencing, core genome phylogenetic analysis, and host-origin prediction by a Bayesian latent class model that incorporated data on the presence of mobile genetic elements (MGEs) among E. coli isolates from multiple vertebrate hosts. A total of 56 multilocus sequence types were identified among the isolates. Five sequence types-ST10, ST38, ST69, ST117, and ST131-were observed among both urine and poultry isolates. Using the Bayesian latent class model, we estimated that 19% (10/52) of the clinical E. coli isolates in our population were foodborne zoonotic strains. These data suggest that a substantial portion of pediatric UTIs in the Washington DC region may be caused by E. coli strains originating in food animals and likely transmitted via contaminated poultry meat.IMPORTANCEEscherichia coli UTIs are a heavy public health burden and can have long-term negative health consequences for pediatric patients. E. coli has an extremely broad host range, including humans, chickens, turkeys, pigs, and cattle. E. coli derived from food animals is a frequent contaminant of retail meat products, but little is known about the risk these strains pose to pediatric populations. Quantifying the proportion of pediatric UTIs caused by food-animal-derived E. coli, characterizing the highest-risk strains, and identifying their primary reservoir species could inform novel intervention strategies to reduce UTI burden in this vulnerable population. Our results suggest that retail poultry meat may be an important vehicle for pediatric exposure to zoonotic E. coli strains capable of causing UTIs. Vaccinating poultry against the highest-risk strains could potentially reduce poultry colonization, poultry meat contamination, and downstream pediatric infections.

Allergic rhinitis (AR) and asthma (AS) are two of the most common chronic respiratory diseases and a major public health concern. Multiple studies have demonstrated the role of the nasal bacteriome in AR and AS, but little is known about the airway mycobiome and its potential association to airway inflammatory diseases. Here we used the internal transcriber spacers (ITS) 1 and 2 and high-throughput sequencing to characterize the nasal mycobiome of 339 individuals with AR, AR with asthma (ARAS), AS and healthy controls (CT). Seven to ten of the 14 most abundant fungal genera (Malassezia, Alternaria, Cladosporium, Penicillium, Wallemia, Rhodotorula, Sporobolomyces, Naganishia, Vishniacozyma, and Filobasidium) in the nasal cavity differed significantly (p ≤ 0.049) between AS, AR or ARAS, and CT. However, none of the same genera varied significantly between the three respiratory disease groups. The nasal mycobiomes of AR and ARAS patients showed the highest intra-group diversity, while CT showed the lowest. Alpha-diversity indices of microbial richness and evenness only varied significantly (p ≤ 0.024) between AR or ARAS and CT, while all disease groups showed significant differences (p ≤ 0.0004) in microbial structure (i.e., beta-diversity indices) when compared to CT samples. Thirty metabolic pathways (PICRUSt2) were differentially abundant (Wald’s test) between AR or ARAS and CT patients, but only three of them associated with 5-aminoimidazole ribonucleotide (AIR) biosynthesis were over abundant (log2 Fold Change >0.75) in the ARAS group. AIR has been associated to fungal pathogenesis in plants. Spiec-Easi fungal networks varied among groups, but AR and ARAS showed more similar interactions among their members than with those in the CT mycobiome; this suggests chronic respiratory allergic diseases may disrupt fungal connectivity in the nasal cavity. This study contributes valuable fungal data and results to understand the relationships between the nasal mycobiome and allergy-related conditions. It demonstrates for the first time that the nasal mycobiota varies during health and allergic rhinitis (with and without comorbid asthma) and reveals specific taxa, metabolic pathways and fungal interactions that may relate to chronic airway disease.

Despite their genetic adaptation to local conditions, plants often achieve ecological success through symbiotic associations with fungal endophytes. However, the habitat-specific functionality of these interactions and their potential to drive plant adaptation to new environments remain uncertain. In this study, we tested this using the vascular flora of the Antarctic tundra (Colobanthus quitensis and Deschampsia antarctica), an extreme environment where fungal endophytes are known for playing important ecological roles. After characterizing the root-associated fungal endophyte communities of both species in two distinct Antarctic terrestrial habitats-hill and coast-we experimentally assessed the contribution of fungal endophytes to plant adaptation in each habitat. The field reciprocal transplant experiment involved removing endophytes from a set of plants and crossing symbiotic status (with and without endophytes) with habitat for both species, aiming to assess plant performance and fitness. The diversity of root fungal endophytes was similar between habitats and mainly explained by plant species, although habitat-specific endophyte community structures were identified in D. antarctica. Endophytes significantly influenced C. quitensis homeostatic regulation, including oxidative stress and osmotic control, as well as plant fitness in both environments. By contrast, the effect of endophytes on D. antarctica was particularly evident in coastal sites, suggesting an endophyte-mediated improvement in local adaptation. Altogether, our results suggest that the two Antarctic vascular plant species follow different strategies in recruiting and developing functional symbiosis with root-associated fungal communities. While C. quitensis is more generalist, D. antarctica establishes specific interactions with habitat-specific microbial symbionts, predominantly in the most stressful environmental context.

Before December 2020, Antarctica had remained free of COVID-19 cases. The main concern during the pandemic was the limited health facilities available at Antarctic stations to deal with the disease as well as the potential impact of SARS-CoV-2 on Antarctic wildlife through reverse zoonosis. In December 2020, 60 cases emerged in Chilean Antarctic stations, disrupting the summer campaign with ongoing isolation needs. The SARS-CoV-2 RNA was detected in the wastewater of several scientific stations. In Antarctica, treated wastewater is discharged directly into the seawater. No studies currently address the recovery of infectious virus particles from treated wastewater, but their presence raises the risk of infecting wildlife and initiating new replication cycles. This study highlights the initial virus detection in wastewater from Antarctic stations, identifying viral RNA via RT-qPCR targeting various genomic regions. The virus’s RNA was found in effluent from two wastewater plants at Maxwell Bay and O’Higgins Station on King George Island and the Antarctic Peninsula, respectively. This study explores the potential for the reverse zoonotic transmission of SARS-CoV-2 from humans to Antarctic wildlife due to the direct release of viral particles into seawater. The implications of such transmission underscore the need for continued vigilance and research.

Microbes play an important role in coastal and estuarine waters. We present 93 metagenomes and 677 metagenome-assembled genomes (MAGs) from Comau Fjord, Patagonia (42°S), to further understand the microbial dynamics and their response to anthropogenic disturbances. These data represent a spatially (35-km transect) and temporally (2016 to 2019) explicit data set.

In 2020, more than 21,000 tons of diesel oil were released accidently into the environment with most of it contaminating water bodies. There is an urgent need for sustainable technologies to clean up rivers and oceans to protect wildlife and human health. One solution is harnessing the power of bacterial consortia; however isolated microbes from different environments have shown low diesel bioremediation rates in seawater thus far. An outstanding question is whether Antarctic microorganisms that thrive in environments polluted with hydro-carbons exhibit better diesel degrading activities when propagated at higher temperatures than those encoun-tered in their natural ecosystems.Here, we isolated bacterial consortia, LR-30 (30 degrees C) and LR-10 (10 degrees C), from the Antarctic rhizosphere soil of Deschampsia antarctica (Livingston Island), that used diesel oil as the only carbon substrate. We found that LR-30 and LR-10 batch bioreactors metabolized nearly the entire diesel content when the initial concentration was 10 (g/L) in seawater. Increasing the initial diesel concentration to 50 gDiesel/L, LR-30 and LR-10 bioconverted 33.4 and 31.2 gDiesel/L in 7 days, respectively. The 16S rRNA gene sequencing profiles revealed that the dominant bacterial genera of the inoculated LR-30 community were Achromobacter (50.6%), Pseudomonas (25%) and Rhodanobacter (14.9%), whereas for LR-10 were Pseudomonas (58%), Candidimonas (10.3%) and Renibacterium (7.8%). We also established continuous bioreactors for diesel biodegradation where LR-30 bioremediated diesel at an unprecedent rate of (34.4 g/L per day), while LR-10 achieved (24.5 g/L per day) at 10 degrees C for one month. The abundance of each bacterial genera present significantly fluctuated at some point during the diesel biore-mediation process, yet Achromobacter and Pseudomonas were the most abundant member at the end of the batch and continuous bioreactors for LR-30 and LR-10, respectively.

Allergic rhinitis and asthma are two of the most common chronic respiratory diseases in developed countries and have become a major public health concern. Substantial evidence has suggested a strong link between respiratory allergy and upper airway dysbacteriosis, but the role of the oral bacteriota is still poorly understood. Here we used 16S rRNA massive parallel sequencing to characterize the oral bacteriome of 344 individuals with allergic rhinitis (AR), allergic rhinitis with asthma (ARAS), asthma (AS) and healthy controls (CT). Four of the most abundant (>2%) phyla (Actinobacteriota, Firmicutes, Fusobacteriota, and Proteobacteria) and 10 of the dominant genera (Actinomyces, Fusobacterium, Gemella, Haemophilus, Leptotrichia, Neisseria, Porphyromonas, Prevotella, Streptococcus, and Veillonella) in the oral cavity differed significantly (p ≤ 0.03) between AR, ARAS or AS and CT groups. The oral bacteriome of ARAS patients showed the highest intra-group diversity, while CT showed the lowest. All alpha-diversity indices of microbial richness and evenness varied significantly (p ≤ 0.022) in ARAS vs. CT and ARAS vs. AR, but they were not significantly different in AR vs. CT. All beta-diversity indices of microbial structure (Unifrac, Bray-Curtis, and Jaccard distances) differed significantly (p ≤ 0.049) between each respiratory disease group and controls. Bacteriomes of AR and ARAS patients showed 15 and 28 upregulated metabolic pathways (PICRUSt2) mainly related to degradation and biosynthesis (p < 0.05). A network analysis (SPIEC-EASI) of AR and ARAS bacteriomes depicted simpler webs of interactions among their members than those observed in the bacteriome of CT, suggesting chronic respiratory allergic diseases may disrupt bacterial connectivity in the oral cavity. This study, therefore, expands our understanding of the relationships between the oral bacteriome and allergy-related conditions. It demonstrates for the first time that the mouth harbors distinct bacteriotas during health and allergic rhinitis (with and without comorbid asthma) and identifies potential taxonomic and functional microbial biomarkers of chronic airway disease.

Abstract. Persistent organic pollutants (POPs) are synthetic compounds that were intentionally produced in large quantities and have been distributed in the global environment, originating a threat due to their persistence, bioaccumulative potential, and toxicity. POPs reach the Antarctic continent through long-range atmospheric transport (LRAT). In these areas, low temperatures play a significant role in the environmental fate of POPs, retaining them for a long time due to cold trapping by diffusion and wet deposition, acting as a net sink for many POPs. However, in the current context of climate change, the remobilization of POPs that were trapped in water, ice, and soil for decades is happening. Therefore, continuous monitoring of POPs in polar air is necessary to assess whether there is a recent re-release of historical pollutants back to the environment. We reviewed the scientific literature on atmospheric levels of several POP families (polychlorinated biphenyls – PCBs, hexachlorobenzene – HCB, hexachlorocyclohexanes – HCHs, and dichlorodiphenyltrichloroethane – DDT) from 1980 to 2021. We estimated the atmospheric half-life using characteristic decreasing times (TD). We observed that HCB levels in the Antarctic atmosphere were higher than the other target organochlorine pesticides (OCPs), but HCB also displayed higher fluctuations and did not show a significant decrease over time. Conversely, the atmospheric levels of HCHs, some DDTs, and PCBs have decreased significantly. The estimated atmospheric half-lives for POPs decreased in the following order: 4,4' DDE (13.5 years) > 4,4' DDD (12.8 years) > 4,4' DDT (7.4 years) > 2,4' DDE (6.4 years) > 2,4' DDT (6.3 years) > α-HCH (6 years) > HCB (6 years) > γ-HCH (4.2 years). For PCB congeners, they decreased in the following order: PCB 153 (7.6 years) > PCB 138 (6.5 years) > PCB 101 (4.7 years) > PCB 180 (4.6 years) > PCB 28 (4 years) > PCB 52 (3.7 years) > PCB 118 (3.6 years). For HCH isomers and PCBs, the Stockholm Convention (SC) ban on POPs did have an impact on decreasing their levels during the last decades. Nevertheless, their ubiquity in the Antarctic atmosphere shows the problematic issues related to highly persistent synthetic chemicals.

Microorganisms are the most diverse life form on the planet and are critical for maintaining the geochemical cycles, especially in extreme environments. Bacterial communities are dynamic and respond directly to changes in abiotic conditions; among these communities, poly-extremophiles are particularly sensitive to perturbations due to their high specialization. Salar de Huasco is a high-altitude wetland located on the Chilean Altiplano exhibiting several conditions considered extreme for life, including negative water balance, extreme variations in temperature and pH values, high UV radiation, and the presence of various toxic metal(oids). However, previous reports have revealed a diverse bacterial community that has adapted to these conditions, here, we aimed to determine whether microbial community diversity and composition changed in response to geographical and seasonal variations. We found that there are significant differences in diversity, abundance, and composition in bacterial taxa that could be attributed to local geographical and seasonal variations, which in turn, can be associated with microbial traits. In conclusion, in this poly-extreme environment, small-scale changes can trigger significant changes in the microbial communities that maintain basic biogeochemical cycles. Further in depth analysis of microbial functionality and geo-ecological dynamics are necessary to better understand the relationships between seasonal changes and bacterial communities.

Most experiments studying bacterial microbiomes rely on the PCR amplification of all or part of the gene for the 16S rRNA subunit, which serves as a biomarker for identifying and quantifying the various taxa present in a microbiome sample. Several computational methods exist for analyzing 16S amplicon sequencing. However, the most-used bioinformatics tools cannot produce high quality genus-level or species-level taxonomic calls and may underestimate the potential accuracy of these calls. We used 16S sequencing data from mock bacterial communities to evaluate the sensitivity and specificity of several bioinformatics pipelines and genomic reference libraries used for microbiome analyses, concentrating on measuring the accuracy of species-level taxonomic assignments of 16S amplicon reads. We evaluated the tools DADA2, QIIME 2, Mothur, PathoScope 2, and Kraken 2 in conjunction with reference libraries from Greengenes, SILVA, Kraken 2, and RefSeq. Profiling tools were compared using publicly available mock community data from several sources, comprising 136 samples with varied species richness and evenness, several different amplified regions within the 16S rRNA gene, and both DNA spike-ins and cDNA from collections of plated cells. PathoScope 2 and Kraken 2, both tools designed for whole-genome metagenomics, outperformed DADA2, QIIME 2 using the DADA2 plugin, and Mothur, which are theoretically specialized for 16S analyses. Evaluations of reference libraries identified the SILVA and RefSeq/Kraken 2 Standard libraries as superior in accuracy compared to Greengenes. These findings support PathoScope and Kraken 2 as fully capable, competitive options for genus- and species-level 16S amplicon sequencing data analysis, whole genome sequencing, and metagenomics data tools.

Viruses are key players in marine environments, affecting food webs and biogeochemical cycles. We present 48 viral metagenomes and 5,656 viral operational taxonomic units (vOTUs) from Comau Fjord, Patagonia (42°S), to understand viral-mediated processes in coastal and estuarine waters. These data represent a spatial (35-km transect, two depths) and seasonal (winter and fall) data set.