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1.
Proc Natl Acad Sci U S A ; 118(43)2021 10 26.
Article in English | MEDLINE | ID: mdl-34667124

ABSTRACT

A high-fat diet (HFD) can induce hyperglycemia and metabolic syndromes that, in turn, can trigger visual impairment. To evaluate the acute effects of HFD feeding on retinal degeneration, we assessed retinal function and morphology, inflammatory state, oxidative stress, and gut microbiome in dystrophic retinal degeneration 10 (rd10) mice, a model of retinitis pigmentosa, fed an HFD for 2 to 3 wk. Short-term HFD feeding impaired retinal responsiveness and visual acuity and enhanced photoreceptor degeneration, microglial cell activation, and Müller cell gliosis. HFD consumption also triggered the expression of inflammatory and oxidative markers in rd10 retinas. Finally, an HFD caused gut microbiome dysbiosis, increasing the abundance of potentially proinflammatory bacteria. Thus, HFD feeding drives the pathological processes of retinal degeneration by promoting oxidative stress and activating inflammatory-related pathways. Our findings suggest that consumption of an HFD could accelerate the progression of the disease in patients with retinal degenerative disorders.


Subject(s)
Diet, High-Fat/adverse effects , Retinal Degeneration/etiology , Retinitis Pigmentosa/etiology , Animals , Cell Death , Disease Models, Animal , Electroretinography , Female , Gastrointestinal Microbiome , Glucose Intolerance , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Models, Biological , Oxidative Stress , Photoreceptor Cells, Vertebrate/pathology , Retina/metabolism , Retina/pathology , Retinal Degeneration/metabolism , Retinal Degeneration/pathology , Retinitis Pigmentosa/metabolism , Retinitis Pigmentosa/pathology
2.
Environ Microbiol ; 25(12): 3151-3160, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37696769

ABSTRACT

Marine viruses play a major role in the energy and nutrient cycle and affect the evolution of their hosts. Despite their importance, there is still little knowledge about RNA viruses. Here, we have explored the Atlantic Ocean, from surface to deep (4.296 m), and used viromics and quantitative methods to unveil the genomics, biogeography, and the mass contribution of RNA viruses to the total viroplankton. A total of 2481 putative RNA viral contigs (>500 bp) and 107 larger bona fide RNA viral genomes (>2.5 kb) were identified; 88 of them representing novel viruses belonging mostly to two clades: Yangshan assemblage (sister clade to the class Alsuviricetes) and Nodaviridae. These viruses were highly endemic and locally abundant, with little or no presence in other oceans since only ≈15% of them were found in at least one of the Tara sampling metatranscriptomes. Quantitative data indicated that the abundance of RNA viruses in the surface and deep chlorophyll maximum zone was within ≈106 VLP/mL representing a potential contribution of 5.2%-24.4% to the total viroplankton community (DNA and RNA viruses), with DNA viruses being the predominant members (≈107 VLP/mL). However, for the deep sample, the observed trend was the opposite, although as further discussed, several biases should be considered. Together these results contribute to our understanding of the diversity, abundance, and distribution of RNA viruses in the oceans and provide a basis for further investigation into their ecological roles and biogeography.


Subject(s)
RNA Viruses , Viruses , Oceans and Seas , RNA Viruses/genetics , Atlantic Ocean , RNA , Seawater
3.
Environ Microbiol ; 23(3): 1584-1593, 2021 03.
Article in English | MEDLINE | ID: mdl-33368907

ABSTRACT

Metagenomics and single-cell genomics have enabled the discovery of relevant uncultured microbes. Recently, single-virus genomics (SVG), although still in an incipient stage, has opened new avenues in viral ecology by allowing the sequencing of one single virus at a time. The investigation of methodological alternatives and optimization of existing procedures for SVG is paramount to deliver high-quality genomic data. We report a sequencing dataset of viral single-amplified genomes (vSAGs) from cultured and uncultured viruses obtained by applying different conditions in each SVG step, from viral preservation and novel whole-genome amplification (WGA) to sequencing platforms and genome assembly. Sequencing data showed that cryopreservation and mild fixation were compatible with WGA, although fresh samples delivered better genome quality data. The novel TruPrime WGA, based on primase-polymerase features, and WGA-X employing a thermostable phi29 polymerase, were proven to be with sufficient sensitivity in SVG. The Oxford Nanopore (ON) sequencing platform did not provide a significant improvement of vSAG assembly compared to Illumina alone. Finally, the SPAdes assembler performed the best. Overall, our results represent a valuable genomic dataset that will help to standardized and advance new tools in viral ecology.


Subject(s)
Genome, Viral , Metagenomics , Genomics , High-Throughput Nucleotide Sequencing
4.
Environ Microbiol ; 23(7): 3477-3498, 2021 07.
Article in English | MEDLINE | ID: mdl-34110059

ABSTRACT

Microbial communities in hypersaline underground waters derive from ancient organisms trapped within the evaporitic salt crystals and are part of the poorly known subterranean biosphere. Here, we characterized the viral and prokaryotic assemblages present in the hypersaline springs that dissolve Triassic-Keuper evaporite rocks and feed the Añana Salt Valley (Araba/Alava, Basque Country, Spain). Four underground water samples (around 23% total salinity) with different levels of exposure to the open air were analysed by means of microscopy and metagenomics. Cells and viruses in the spring water had lower concentrations than what are normally found in hypersaline environments and seemed to be mostly inactive. Upon exposure to the open air, there was an increase in activity of both cells and viruses as well as a selection of phylotypes. The underground water was inhabited by a rich community harbouring a diverse set of genes coding for retinal binding proteins. A total of 35 viral contigs from 15 to 104 kb, representing partial or total viral genomes, were assembled and their evolutionary changes through the spring system were followed by SNP analysis and metagenomic island tracking. Overall, both the viral and the prokaryotic assemblages changed quickly upon exposure to the open air conditions.


Subject(s)
Metagenomics , Viruses , Metagenome/genetics , Phylogeny , Salinity , Viruses/genetics
5.
Environ Microbiol ; 22(5): 1748-1763, 2020 05.
Article in English | MEDLINE | ID: mdl-31840364

ABSTRACT

Members of the SAR11 clade, despite their high abundance, are often poorly represented by metagenome-assembled genomes. This fact has hampered our knowledge about their ecology and genetic diversity. Here we examined 175 SAR11 genomes, including 47 new single-amplified genomes. The presence of the first genomes associated with subclade IV suggests that, in the same way as subclade V, they might be outside the proposed Pelagibacterales order. An expanded phylogenomic classification together with patterns of metagenomic recruitment at a global scale have allowed us to define new ecogenomic units of classification (genomospecies), appearing at different, and sometimes restricted, metagenomic data sets. We detected greater microdiversity across the water column at a single location than in samples collected from similar depth across the global ocean, suggesting little influence of biogeography. In addition, pangenome analysis revealed that the flexible genome was essential to shape genomospecies distribution. In one genomospecies preferentially found within the Mediterranean, a set of genes involved in phosphonate utilization was detected. While another, with a more cosmopolitan distribution, was unique in having an aerobic purine degradation pathway. Together, these results provide a glimpse of the enormous genomic diversity within this clade at a finer resolution than the currently defined clades.


Subject(s)
Genome, Bacterial/genetics , Hyphomicrobiaceae/genetics , Genomics , Hyphomicrobiaceae/classification , Mediterranean Region , Metagenome/genetics , Metagenomics , Oceans and Seas , Organophosphonates/metabolism , Phylogeny , Purines/metabolism , Seawater/microbiology , Water Microbiology
6.
Environ Microbiol ; 21(12): 4582-4596, 2019 12.
Article in English | MEDLINE | ID: mdl-31342640

ABSTRACT

Wastewater treatment plants effluents are considered as hotspots for the dispersion of antibiotic resistance genes (ARGs) into natural ecosystems. The bacterial resistome (ARG collection in a metagenome) analyses have provided clues on antibacterial resistance dynamics. However, viruses and vesicles are frequently ignored. Here, we addressed the bacterial, viral and vesicle resistomes from a representative wastewater effluent in natural conditions and amended with polymyxin, which is used as a last resort antibiotic. Metagenomics showed that the natural prokaryotic resistome was vast (9000 ARG hits/Gb metagenome) and diverse, while viral resistome was two orders of magnitude lower (50 ARG hits/Gb metagenome) suggesting that viruses rarely encoded ARGs. After polymyxin amendment, data showed no ARG enrichment - including to polymyxin - in the microbiome. Remarkably, microbiomes responded to polymyxin with a vast release of putative vesicles (threefold increase compared with the control), which might be used as 'traps' to decrease the antibiotic concentration. Intriguingly, although polymyxin resistance genes (PRGs) were rare in the microbiome (0.018% of total ARG found), in the viral and vesicle fractions, PRGs were more abundant (0.5%-0.8% of total ARG found). Our data suggest that vesicles could have a more active role in the context of transmission of antibiotic resistances.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Microbial , Extracellular Vesicles , Microbiota/drug effects , Wastewater/microbiology , Water Microbiology , Bacteria/drug effects , Bacteria/genetics , Genes, Bacterial , Metagenome , Viruses/genetics
7.
Environ Microbiol ; 21(6): 2029-2042, 2019 06.
Article in English | MEDLINE | ID: mdl-30724439

ABSTRACT

Salar de Uyuni (SdU) is the largest hypersaline salt flat and the highest lithium reservoir on Earth. In addition to extreme temperatures and high UV irradiance, SdU has high concentrations of chaotropic salts which can be important factors in controlling microbial diversity. Here, for the first time we characterize the viral diversity of this hypersaline environment during the two seasons, as well as the physicochemical characteristics and the prokaryotic communities of the analysed samples. Most of the selected samples showed a peculiar physicochemical composition and prokaryotic diversity, mostly different from each other even for samples from locations in close proximity or the same season. In contrast to most hypersaline systems Bacteria frequently outnumbered Archaea. Furthermore, an outstanding percentage of members of Salinibacter sp., likely a species different from the cosmopolitan Salinibacter ruber, was obtained in most of the samples. Viral communities displayed the morphologies normally found in hypersaline environments. Two seasonal samples were chosen for a detailed metagenomic analysis of the viral assemblage. Both viral communities shared common sequences but were dominated by sample-specific viruses, mirroring the differences also observed in physicochemical and prokaryotic community composition. These metaviromes were distinct from those detected in other hypersaline systems analysed to date.


Subject(s)
Archaea/isolation & purification , Bacteria/isolation & purification , Lakes/microbiology , Lakes/virology , Viruses/isolation & purification , Archaea/classification , Archaea/genetics , Bacteria/classification , Bacteria/genetics , Biodiversity , Lakes/analysis , Metagenome , Phylogeny , Salinity , Viruses/classification , Viruses/genetics
8.
Environ Microbiol ; 21(10): 3577-3600, 2019 10.
Article in English | MEDLINE | ID: mdl-31087616

ABSTRACT

Peñahueca is an athalassohaline hypersaline inland ephemeral lake originated under semiarid conditions in the central Iberian Peninsula (Spain). Its chemical composition makes it extreme for microbial life as well as a terrestrial analogue of other planetary environments. To investigate the persistence of microbial life associated with sulfate-rich crusts, we applied cultivation-independent methods (optical and electron microscopy, 16S rRNA gene profiling and metagenomics) to describe the prokaryotic community and its associated viruses. The diversity for Bacteria was very low and was vastly dominated by endospore formers related to Pontibacillus marinus of the Firmicutes phylum. The archaeal assemblage was more diverse and included taxa related to those normally found in hypersaline environments. Several 'metagenome assembled genomes' were recovered, corresponding to new species of Pontibacillus, several species from the Halobacteria and one new member of the Nanohaloarchaeota. The viral assemblage, although composed of the morphotypes typical of high salt systems, showed little similarity to previously isolated/reconstructed halophages. Several putative prophages of Pontibacillus and haloarchaeal hosts were identified. Remarkably, the Peñahueca sulfate-rich metagenome contained CRISPR-associated proteins and repetitions which were over 10-fold higher than in most hypersaline systems analysed so far.


Subject(s)
Archaea/genetics , Bacteria/genetics , Lakes/microbiology , Viruses/genetics , Archaea/classification , Bacteria/classification , Biodiversity , Exobiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Sodium Chloride/metabolism , Spain , Sulfates/metabolism , Viruses/classification
9.
Environ Microbiol ; 18(12): 4564-4578, 2016 12.
Article in English | MEDLINE | ID: mdl-27690185

ABSTRACT

Corals are known to contain a diverse microbiota that plays a paramount role in the physiology and health of holobiont. However, few studies have addressed the variability of bacterial communities within the coral host. In this study, bacterial community composition from the mucus, tissue and skeleton of the scleractinian coral Oculina patagonica were investigated seasonally at two locations in the Western Mediterranean Sea, to further understand how environmental conditions and the coral microbiome structure are related. We used denaturing gradient gel electrophoresis in combination with next-generation sequencing and electron microscopy to characterize the bacterial community. The bacterial communities were significantly different among coral compartments, and coral tissue displayed the greatest changes related to environmental conditions and coral health status. Species belonging to the Rhodobacteraceae and Vibrionaceae families form part of O. patagonica tissues core microbiome and may play significant roles in the nitrogen cycle. Furthermore, sequences related to the coral pathogens, Vibrio mediterranei and Vibrio coralliilyticus, were detected not only in bleached corals but also in healthy ones, even during cold months. This fact opens a new view onto unveiling the role of pathogens in the development of coral diseases in the future.


Subject(s)
Anthozoa/microbiology , Animals , Denaturing Gradient Gel Electrophoresis , Ecosystem , Mediterranean Sea , Vibrio/isolation & purification , Vibrio/physiology
10.
Proc Natl Acad Sci U S A ; 110(28): 11463-8, 2013 Jul 09.
Article in English | MEDLINE | ID: mdl-23801761

ABSTRACT

Planktonic bacteria dominate surface ocean biomass and influence global biogeochemical processes, but remain poorly characterized owing to difficulties in cultivation. Using large-scale single cell genomics, we obtained insight into the genome content and biogeography of many bacterial lineages inhabiting the surface ocean. We found that, compared with existing cultures, natural bacterioplankton have smaller genomes, fewer gene duplications, and are depleted in guanine and cytosine, noncoding nucleotides, and genes encoding transcription, signal transduction, and noncytoplasmic proteins. These findings provide strong evidence that genome streamlining and oligotrophy are prevalent features among diverse, free-living bacterioplankton, whereas existing laboratory cultures consist primarily of copiotrophs. The apparent ubiquity of metabolic specialization and mixotrophy, as predicted from single cell genomes, also may contribute to the difficulty in bacterioplankton cultivation. Using metagenome fragment recruitment against single cell genomes, we show that the global distribution of surface ocean bacterioplankton correlates with temperature and latitude and is not limited by dispersal at the time scales required for nucleotide substitution to exceed the current operational definition of bacterial species. Single cell genomes with highly similar small subunit rRNA gene sequences exhibited significant genomic and biogeographic variability, highlighting challenges in the interpretation of individual gene surveys and metagenome assemblies in environmental microbiology. Our study demonstrates the utility of single cell genomics for gaining an improved understanding of the composition and dynamics of natural microbial assemblages.


Subject(s)
Bacteria/classification , Genome, Bacterial , Marine Biology , Plankton/classification , Water Microbiology , Bacteria/genetics , Geography , Oceans and Seas , Plankton/genetics
11.
Environ Microbiol ; 17(2): 514-26, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25387432

ABSTRACT

Microbial opsin (i.e. retinal-binding protein) dynamics has been studied along a salinity gradient in Santa Pola solar salterns (Alicante, Spain) by using culture-independent approaches and statistical analyses. Five ponds of salinities ranging from 18% to above 40% were sampled nine times along a year. Forty-three opsin-like sequences were retrieved by denaturing gradient gel electrophoresis and clustered into 18 different phylogroups, indicating that their diversity was higher than expected according to previous data. Moreover, the statistical correlation between environmental factors controlling microbial community structure and dynamics of environmental rhodopsin proteins indicated almost identical temporal fluctuations between the opsin-related sequences and their corresponding putative 'producers' in nature. Although most sequences were related to others previously detected in hypersaline environments, some pond-specific opsins putatively belonged to previously uncharacterized hosts. Furthermore, we propose that subtle changes in the bacteriorhodopsin 'retinal proton binding pocket', which is key in the photocycle function, could be the molecular basis behind a fine 'photocycle-tuning' mechanism to avoid inter/intraspecies light-competition in hypersaline environments.


Subject(s)
Archaea/genetics , Bacteria/genetics , Bacteriorhodopsins/chemistry , Rhodopsin/chemistry , Amino Acid Sequence , Archaea/metabolism , Bacteria/metabolism , Bacteriorhodopsins/genetics , Denaturing Gradient Gel Electrophoresis , Rhodopsin/genetics , Salinity , Spain , Water Microbiology
12.
Appl Environ Microbiol ; 81(24): 8445-56, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26431969

ABSTRACT

Comparative genomics, metagenomics, and single-cell technologies have shown that populations of microbial species encompass assemblages of closely related strains. This raises the question of whether individual bacterial lineages respond to the presence of their close relatives by modifying their gene expression or, instead, whether assemblages simply act as the arithmetic addition of their individual components. Here, we took advantage of transcriptome sequencing to address this question. For this, we analyzed the transcriptomes of two closely related strains of the extremely halophilic bacterium Salinibacter ruber grown axenically and in coculture. These organisms dominate bacterial assemblages in hypersaline environments worldwide. The strains used here cooccurred in the natural environment and are 100% identical in their 16S rRNA genes, and each strain harbors an accessory genome representing 10% of its complete genome. Overall, transcriptomic patterns from pure cultures were very similar for both strains. Expression was detected along practically the whole genome albeit with some genes at low levels. A subset of genes was very highly expressed in both strains, including genes coding for the light-driven proton pump xanthorhodopsin, genes involved in the stress response, and genes coding for transcriptional regulators. Expression differences between pure cultures affected mainly genes involved in environmental sensing. When the strains were grown in coculture, there was a modest but significant change in their individual transcription patterns compared to those in pure culture. Each strain sensed the presence of the other and responded in a specific manner, which points to fine intraspecific transcriptomic modulation.


Subject(s)
Bacteroidetes/classification , Bacteroidetes/genetics , High-Throughput Nucleotide Sequencing , Microbial Interactions/genetics , Transcriptome/genetics , Bacterial Proteins/genetics , Bacteroidetes/metabolism , Base Sequence , Coculture Techniques , DNA, Bacterial/genetics , Genome, Bacterial/genetics , Metagenomics , RNA, Ribosomal, 16S/genetics , Rhodopsins, Microbial/genetics , Sequence Analysis, DNA , Transcription, Genetic
13.
Ginecol Obstet Mex ; 83(11): 735-42, 2015 Nov.
Article in Spanish | MEDLINE | ID: mdl-27311172

ABSTRACT

BACKGROUND: Pheochromocytoma is a tumor originating in the chromaffin cells. These tumors secrete catecholamines which act on target organs and cause hypertensive crises. They are rare during pregnancy, and a differential diagnosis must be carried out mainly with pregnancy-induced hypertension. CLINIC CASE: A 22-year-old patient in week 11.5 of pregnancy presented at the Hospital General Dr. Miguel Silva in Morelia, Michoacán with hypertension that had existed for more than two years with poor adherence to treatment. At the time of referral to our unit she presented a hypertensive crisis that had been unresponsive to all antihypertensive treatments. Following the establishment of a study protocol, a diagnosis of posterior left adrenal pheochromocytoma was made. After subsequent pharmacological treatment with alpha and beta blockers, a left adrenalectomy was performed. The patient did not require antihypertensive treatment following surgery. The histopathological report indicates the presence of a nodular and vascularized left adrenal gland weighing 25 g and measuring 5 x 4 x 3.5 cm. Gland consistency was soft, and the presence of cystic lesions in parenchymal tissue was noted. A yellowish-brown color and sponge-like appearance were also present. Such features support a diagnosis of pheochromocytoma. The pregnancy continued normally until week 32, when the patient presented to our department with active-phase preterm labor. A Cesarean section was performed with the delivery of a live female weighing 1400 gr. The infant is currently alive and well. CONCLUSION: Thus, it is important that obstetrician knows this disease and its management during pregnancy; so we present this case report.


Subject(s)
Adrenal Gland Neoplasms/diagnosis , Pheochromocytoma/diagnosis , Pregnancy Complications, Neoplastic/diagnosis , Adrenal Gland Neoplasms/complications , Adrenal Gland Neoplasms/surgery , Adrenalectomy/methods , Diagnosis, Differential , Female , Humans , Hypertension, Pregnancy-Induced/etiology , Pheochromocytoma/complications , Pheochromocytoma/surgery , Pregnancy , Pregnancy Complications, Neoplastic/pathology , Pregnancy Complications, Neoplastic/surgery , Pregnancy Outcome , Young Adult
14.
Environ Microbiol ; 16(9): 2682-98, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24118837

ABSTRACT

Little is known about the diversity and structuring of freshwater microbial communities beyond the patterns revealed by tracing their distribution in the landscape with common taxonomic markers such as the ribosomal RNA. To address this gap in knowledge, metagenomes from temperate lakes were compared to selected marine metagenomes. Taxonomic analyses of rRNA genes in these freshwater metagenomes confirm the previously reported dominance of a limited subset of uncultured lineages of freshwater bacteria, whereas Archaea were rare. Diversification into marine and freshwater microbial lineages was also reflected in phylogenies of functional genes, and there were also significant differences in functional beta-diversity. The pathways and functions that accounted for these differences are involved in osmoregulation, active transport, carbohydrate and amino acid metabolism. Moreover, predicted genes orthologous to active transporters and recalcitrant organic matter degradation were more common in microbial genomes from oligotrophic versus eutrophic lakes. This comparative metagenomic analysis allowed us to formulate a general hypothesis that oceanic- compared with freshwater-dwelling microorganisms, invest more in metabolism of amino acids and that strategies of carbohydrate metabolism differ significantly between marine and freshwater microbial communities.


Subject(s)
Bacteria/classification , Fresh Water/microbiology , Metagenomics , Phylogeny , Salinity , Archaea/classification , Archaea/genetics , Bacteria/genetics , Data Mining , Lakes/microbiology , Metabolic Networks and Pathways , Metagenome , Molecular Sequence Annotation , RNA, Ribosomal/genetics , Seawater/microbiology , Sequence Analysis, DNA
15.
Int J Syst Evol Microbiol ; 64(Pt 8): 2841-2848, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24871776

ABSTRACT

A Gram-reaction-positive bacterial isolate, designated Tü 6233(T), with rudimentary, coral-pink vegetative mycelium that formed neither aerial mycelium nor spores, was isolated from a Brazilian soil sample. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. Cell-wall hydrolysates contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose as the diagnostic sugar. The major fatty acids were iso-C(16 : 0), iso-C(15 : 0) and C(17 : 1)ω8c and the predominant menaquinone was MK-9(H4). The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylinositol, an unknown glycophospholipid and an unknown phospholipid. The DNA G+C content of the strain was 75.4 mol%. The 16S rRNA gene sequence identity with members of the genus Geodermatophilus was 94.2-98.7%. Based on phenotypic, chemotaxonomic and phylogenetic data, strain Tü 6233(T) is proposed to represent a novel species, Geodermatophilus brasiliensis sp. nov., with the type strain Tü 6233(T) ( = DSM 44526(T) = CECT 8402(T)).


Subject(s)
Actinomycetales/classification , Phylogeny , Soil Microbiology , Actinomycetales/genetics , Actinomycetales/isolation & purification , Bacterial Typing Techniques , Base Composition , Brazil , DNA, Bacterial/genetics , Diaminopimelic Acid/chemistry , Fatty Acids/chemistry , Molecular Sequence Data , Nucleic Acid Hybridization , Phospholipids/chemistry , Pigmentation , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistry
16.
Antonie Van Leeuwenhoek ; 106(4): 675-92, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25064091

ABSTRACT

Prokaryotes in the superficial sediments are ecologically important microorganisms that are responsible for the decomposition, mineralization and subsequent recycling of organic matter. The aim of this study was to explore the phylogenetic and functional diversity of halophilic archaea and bacteria isolated from the superficial sediments of solar salterns at Sfax, Tunisia. Sixty four strains were isolated from crystallizer (TS18) and non-crystallizer (M1) ponds and submitted to genotypic characterization and evaluation by amplified ribosomal RNA restriction analysis (ARDRA) techniques. Our findings revealed that the archaeal diversity observed for 29 isolates generated five distinct patterns from the non-crystallizer M1 pond, with Halorubrum chaoviator as the most prevalent cultivable species. However, in the TS18 crystallizer pond, ten restriction patterns were observed, with the prevalence of haloarchaea EB27K, a not yet identified genotype. The construction of a neighbour-joining tree of 16S rRNA gene sequences resulted in the division of the potential new species into two major groups, with four strains closely related to the sequence of the unculturable haloarchaeon EB27K and one strain to the recently described Halovenus aranensis strain. The 35 bacterial strains observed in this work were present only in the non-crystallizer pond (M1) and presented two distinct ARDRA patterns. These strains belonged to the γ-proteobacteria subdivision, with members of Salicola marasensis (83%) being the most predominant species among the isolates. 16S rRNA gene sequencing revealed that Salicola strains displayed different degrees of homogeneity. The results from pulsed field gel electrophoresis assays showed that the Salicola isolates could be clustered in two distinct groups with different genome sizes.


Subject(s)
Archaea/classification , Archaea/isolation & purification , Bacteria/classification , Bacteria/isolation & purification , Geologic Sediments/microbiology , Archaea/genetics , Archaea/metabolism , Bacteria/genetics , Bacteria/metabolism , Cluster Analysis , DNA, Archaeal/chemistry , DNA, Archaeal/genetics , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Seasons , Sequence Analysis, DNA , Tunisia
17.
Environ Microbiome ; 19(1): 16, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38491508

ABSTRACT

BACKGROUND: Anthropogenic activities significantly contribute to the dissemination of antibiotic resistance genes (ARGs), posing a substantial threat to humankind. The development of methods that allow robust ARG surveillance is a long-standing challenge. Here, we use city-scale monitoring of ARGs by using two of the most promising cutting-edge technologies, digital PCR (dPCR) and metagenomics. METHODS: ARG hot-spots were sampled from the urban water and wastewater distribution systems. Metagenomics was used to provide a broad view of ARG relative abundance and richness in the prokaryotic and viral fractions. From the city-core ARGs in all samples, the worldwide dispersed sul2 and tetW conferring resistance to sulfonamide and tetracycline, respectively, were monitored by dPCR and metagenomics. RESULTS: The largest relative overall ARG abundance and richness were detected in the hospital wastewater and the WWTP inlet (up to ≈6,000 ARGs/Gb metagenome) with a large fraction of unclassified resistant bacteria. The abundance of ARGs in DNA and RNA contigs classified as viruses was notably lower, demonstrating a reduction of up to three orders of magnitude compared to contigs associated to prokaryotes. By metagenomics and dPCR, a similar abundance tendency of sul2 and tetW was obtained, with higher abundances in hospital wastewater and WWTP input (≈125-225 ARGs/Gb metagenome). dPCR absolute abundances were between 6,000 and 18,600 copies per ng of sewage DNA (≈105-7 copies/mL) and 6.8 copies/mL in seawater near the WWTP discharging point. CONCLUSIONS: dPCR was more sensitive and accurate, while metagenomics provided broader coverage of ARG detection. While desirable, a reliable correlation of dPCR absolute abundance units into metagenomic relative abundance units was not obtained here (r2 < 0.4) suggesting methodological factors that introduce variability. Evolutionary pressure does not significantly select the targeted ARGs in natural aquatic environments.

18.
Environ Microbiol Rep ; 16(3): e13306, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38923122

ABSTRACT

Human activities are a significant contributor to the spread of antibiotic resistance genes (ARGs), which pose a serious threat to human health. These ARGs can be transmitted through various pathways, including air, within the context of One Health. This study used metagenomics to monitor the resistomes in urban air from two critical locations: a wastewater treatment plant and a hospital, both indoor and outdoor. The presence of cell-like structures was confirmed through fluorescence microscopy. The metagenomic analysis revealed a wide variety of ARGs and a high diversity of antibiotic-resistant bacteria in the airborne particles collected. The wastewater treatment plant showed higher relative abundances with 32 ARG hits per Gb and m3, followed by the main entrance of the hospital (indoor) with ≈5 ARG hits per Gb and m3. The hospital entrance exhibited the highest ARG richness, with a total of 152 different ARGs classified into nine categories of antibiotic resistance. Common commensal and pathogenic bacteria carrying ARGs, such as Moraxella, Staphylococcus and Micrococcus, were detected in the indoor airborne particles of the hospital. Interestingly, no ARGs were shared among all the samples analysed, indicating a highly variable dynamic of airborne resistomes. Furthermore, the study found no ARGs in the airborne viral fractions analysed, suggesting that airborne viruses play a negligible role in the dissemination of ARGs.


Subject(s)
Air Microbiology , Bacteria , Drug Resistance, Bacterial , Metagenomics , Bacteria/genetics , Bacteria/classification , Bacteria/isolation & purification , Bacteria/drug effects , Humans , Drug Resistance, Bacterial/genetics , One Health , Metagenome , Wastewater/microbiology , Genes, Bacterial/genetics , Hospitals , Anti-Bacterial Agents/pharmacology , Cities
19.
mSphere ; 9(5): e0007324, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38666797

ABSTRACT

The signs of climate change are undeniable, and the impact of these changes on ecosystem function heavily depends on the response of microbes that underpin the food web. Antarctic ice shelf is a massive mass of floating ice that extends from the continent into the ocean, exerting a profound influence on global carbon cycles. Beneath Antarctic ice shelves, marine ice stores valuable genetic information, where marine microbial communities before the industrial revolution are archived. Here, in this proof-of-concept, by employing a combination of single-cell technologiesand metagenomics, we have been able to sequence frozen microbial DNA (≈300 years old) stored in the marine ice core B15 collected from the Filchnner-Ronne Ice Shelf. Metagenomic data indicated that Proteobacteria and Thaumarchaeota (e.g., Nitrosopumilus spp.), followed by Actinobacteria (e.g., Actinomarinales), were abundant. Remarkably, our data allow us to "travel to the past" and calibrate genomic and genetic evolutionary changes for ecologically relevant microbes and functions, such as Nitrosopumilus spp., preserved in the marine ice (≈300 years old) with those collected recently in seawater under an ice shelf (year 2017). The evolutionary divergence for the ammonia monooxygenase gene amoA involved in chemolithoautotrophy was about 0.88 amino acid and 2.8 nucleotide substitution rate per 100 sites in a century, while the accumulated rate of genomic SNPs was 2,467 per 1 Mb of genome and 100 years. Whether these evolutionary changes remained constant over the last 300 years or accelerated during post-industrial periods remains an open question that will be further elucidated. IMPORTANCE: Several efforts have been undertaken to predict the response of microbes under climate change, mainly based on short-term microcosm experiments under forced conditions. A common concern is that manipulative experiments cannot properly simulate the response of microbes to climate change, which is a long-term evolutionary process. In this proof-of-concept study with a limited sample size, we demonstrate a novel approach yet to be fully explored in science for accessing genetic information from putative past marine microbes preserved under Antarctic ice shelves before the industrial revolution. This potentially allows us estimating evolutionary changes as exemplified in our study. We advocate for gathering a more comprehensive Antarctic marine ice core data sets across various periods and sites. Such a data set would enable the establishment of a robust baseline, facilitating a better assessment of the potential effects of climate change on key genetic signatures of microbes.


Subject(s)
Bacteria , Climate Change , Ice Cover , Metagenomics , Microbiota , Seawater , Antarctic Regions , Ice Cover/microbiology , Microbiota/genetics , Metagenomics/methods , Bacteria/genetics , Bacteria/classification , Seawater/microbiology , Archaea/genetics , Archaea/classification , Ecosystem , Single-Cell Analysis , Phylogeny
20.
Nat Commun ; 14(1): 8295, 2023 Dec 14.
Article in English | MEDLINE | ID: mdl-38097581

ABSTRACT

Viruses play an important role in the marine ecosystem. However, our comprehension of viruses inhabiting the dark ocean, and in particular, under the Antarctic Ice Shelves, remains limited. Here, we mine single-cell genomic, transcriptomic, and metagenomic data to uncover the viral diversity, biogeography, activity, and their role as metabolic facilitators of microbes beneath the Ross Ice Shelf. This is the largest Antarctic ice shelf with a major impact on global carbon cycle. The viral community found in the cavity under the ice shelf mainly comprises endemic viruses adapted to polar and mesopelagic environments. The low abundance of genes related to lysogenic lifestyle (<3%) does not support a predominance of the Piggyback-the-Winner hypothesis, consistent with a low-productivity habitat. Our results indicate a viral community actively infecting key ammonium and sulfur-oxidizing chemolithoautotrophs (e.g. Nitrosopumilus spp, Thioglobus spp.), supporting a "kill-the-winner" dynamic. Based on genome analysis, these viruses carry specific auxiliary metabolic genes potentially involved in nitrogen, sulfur, and phosphorus acquisition. Altogether, the viruses under Antarctic ice shelves are putatively involved in programming the metabolism of ecologically relevant microbes that maintain primary production in these chemosynthetically-driven ecosystems, which have a major role in global nutrient cycles.


Subject(s)
Ecosystem , Viruses , Antarctic Regions , Archaea , Viruses/genetics , Sulfur , Ice Cover
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