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Have you ever sought to use metagenomic DNA sequences reported in scientific publications? Were you successful? Here, we reveal that metagenomes from no fewer than 20% of the papers found in our literature search, published between 2016 and 2019, were not deposited in a repository or were simply inaccessible. The proportion of inaccessible data within the literature has been increasing year-on-year. Noncompliance with Open Data is best predicted by the scientific discipline of the journal. The number of citations, journal type (e.g., Open Access or subscription journals), and publisher are not good predictors of data accessibility. However, many publications in high-impact factor journals do display a higher likelihood of accessible metagenomic data sets. Twenty-first century science demands compliance with the ethical standard of data sharing of metagenomes and DNA sequence data more broadly. Data accessibility must become one of the routine and mandatory components of manuscript submissions-a requirement that should be applicable across the increasing number of disciplines using metagenomics. Compliance must be ensured and reinforced by funders, publishers, editors, reviewers, and, ultimately, the authors.
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Acceso a la Información , Metagenoma , Publicaciones/estadística & datos numéricos , Bibliometría , Factor de Impacto de la Revista , Publicación de Acceso AbiertoRESUMEN
This study shows that Escherichia coli can be temporarily enriched in zooplankton under natural conditions and that these bacteria can belong to different phylogroups and sequence types (STs), including environmental, clinical, and animal isolates. We isolated 10 E. coli strains and sequenced the genomes of two of them. Phylogenetically, the two isolates were closer to strains isolated from poultry meat than to freshwater E. coli, albeit their genomes were smaller than those of the poultry isolates. After isolation and fluorescent protein tagging of strains ED1 and ED157, we show that Daphnia sp. can take up these strains and release them alive again, thus becoming a temporary host for E. coli. In a chemostat experiment, we show that this association does not prolong bacterial long-term survival, but at low abundances it also does not significantly reduce bacterial numbers. We demonstrate that E. coli does not belong to the core microbiota of Daphnia, suffers from competition by the natural Daphnia microbiota, but can profit from its carapax to survive in water. All in all, this study suggests that the association of E. coli with Daphnia is only temporary, but the cells are viable therein, and this might allow encounters with other bacteria for genetic exchange and potential genomic adaptation to the freshwater environment. IMPORTANCE The contamination of freshwater with feces-derived bacteria is a major concern regarding drinking water acquisition and recreational activities. Ecological interactions promoting their persistence are still very scarcely studied. This study, which analyses the survival of E. coli in the presence of zooplankton, is thus of ecological and water safety relevance.
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Agua Potable , Escherichia coli , Animales , Bacterias , Daphnia/microbiología , Escherichia coli/genética , Heces/microbiología , Agua Dulce/microbiología , Zooplancton/microbiologíaRESUMEN
Infectious agents such as the bacteria Vibrio aestuarianus or Ostreid herpesvirus 1 have been repeatedly associated with dramatic disease outbreaks of Crassostrea gigas beds in Europe. Beside roles played by these pathogens, microbial infections in C. gigas may derive from the contribution of a larger number of microorganisms than previously thought, according to an emerging view supporting the polymicrobial nature of bivalve diseases. In this study, the microbial communities associated with a large number of C. gigas samples collected during recurrent mortality episodes at different European sites were investigated by real-time PCR and 16SrRNA gene-based microbial profiling. A new target enrichment next-generation sequencing protocol for selective capturing of 884 phylogenetic and virulence markers of the potential microbial pathogenic community in oyster tissue was developed allowing high taxonomic resolution analysis of the bivalve pathobiota. Comparative analysis of contrasting C. gigas samples conducted using these methods revealed that oyster experiencing mortality outbreaks displayed signs of microbiota disruption associated with the presence of previously undetected potential pathogenic microbial species mostly belonging to genus Vibrio and Arcobacter. The role of these species and their consortia should be targeted by future studies aiming to shed light on mechanisms underlying polymicrobial infections in C. gigas.
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Bacterias/aislamiento & purificación , Crassostrea/microbiología , Microbiota , Animales , Bacterias/clasificación , Bacterias/genética , Virus ADN/clasificación , Virus ADN/genética , Virus ADN/aislamiento & purificación , Europa (Continente) , Secuenciación de Nucleótidos de Alto Rendimiento , Microbiota/genética , Tipificación Molecular , Filogenia , ARN Bacteriano , ARN Ribosómico 16S , Reacción en Cadena en Tiempo Real de la Polimerasa , Vibrio/genética , Vibrio/aislamiento & purificación , Virulencia/genéticaRESUMEN
Notwithstanding the fundamental role that environmental microbes play for ecosystem functioning, data on how microbes react to disturbances are still scarce, and most factors that confer stability to microbial communities are unknown. In this context, antibiotic discharge into the environment is considered a worldwide threat for ecosystems with potential risks to human health. We therefore tested resilience of microbial communities challenged by the presence of an antibiotic. In a continuous culture experiment, we compared the abundance, composition and diversity of microbial communities undisturbed or disturbed by the constant addiction of tetracycline in low (10 µg/L) or intermediate (100 µg/L) concentration (press disturbance). Further, the bacterial communities in the three treatments had to face the sudden pulse disturbance of adding an allochthonous bacterium (Escherichia coli). Tetracycline, even at low concentrations, affected microbial communities by changing their phylogenetic composition and causing cell aggregation. This, however, did not coincide with a reduced microbial diversity, but was mainly caused by a shift in dominance of specific bacterial families. Moreover, the less disturbed community (10 µg/L tetracycline) was sometimes more similar to the control and sometimes more similar to heavily disturbed community (100 µg/L tetracycline). All in all, we could not see a pattern where the communities disturbed with antibiotics were less resilient to a second disturbance introducing E. coli, but they seemed to be able to buffer the input of the allochthonous strain in a similar manner as the control.
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Bacterias/efectos de los fármacos , Ecosistema , Microbiota/genética , Filogenia , Antibacterianos/efectos adversos , Bacterias/genética , Biodiversidad , Cadena Alimentaria , ARN Ribosómico 16S/genéticaRESUMEN
BACKGROUND: Many cyanobacteria are capable of fixing atmospheric nitrogen, playing a crucial role in biogeochemical cycling. Little is known about freshwater unicellular cyanobacteria Synechococcus spp. at the genomic level, despite being recognised of considerable ecological importance in aquatic ecosystems. So far, it has not been shown whether these unicellular picocyanobacteria have the potential for nitrogen fixation. Here, we present the draft-genome of the new pink-pigmented Synechococcus-like strain Vulcanococcus limneticus. sp. nov., isolated from the volcanic Lake Albano (Central Italy). RESULTS: The novel species Vulcanococcus limneticus sp. nov. falls inside the sub-cluster 5.2, close to the estuarine/marine strains in a maximum-likelihood phylogenetic tree generated with 259 marker genes with representatives from marine, brackish, euryhaline and freshwater habitats. V.limneticus sp. nov. possesses a complete nitrogenase and nif operon. In an experimental setup under nitrogen limiting and non-limiting conditions, growth was observed in both cases. However, the nitrogenase genes (nifHDK) were not transcribed, i.e., V.limneticus sp. nov. did not fix nitrogen, but instead degraded the phycobilisomes to produce sufficient amounts of ammonia. Moreover, the strain encoded many other pathways to incorporate ammonia, nitrate and sulphate, which are energetically less expensive for the cell than fixing nitrogen. The association of the nif operon to a genomic island, the relatively high amount of mobile genetic elements (52 transposases) and the lower observed GC content of V.limneticus sp. nov. nif operon (60.54%) compared to the average of the strain (68.35%) support the theory that this planktonic strain may have obtained, at some point of its evolution, the nif operon by horizontal gene transfer (HGT) from a filamentous or heterocystous cyanobacterium. CONCLUSIONS: In this study, we describe the novel species Vulcanococcus limneticus sp. nov., which possesses a complete nif operon for nitrogen fixation. The finding that in our experimental conditions V.limneticus sp. nov. did not express the nifHDK genes led us to reconsider the actual ecological meaning of these accessory genes located in genomic island that have possibly been acquired via HGT.
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Cianobacterias/genética , Transferencia de Gen Horizontal , Nitrógeno/metabolismo , Nitrogenasa/genética , Operón , Cianobacterias/clasificación , Cianobacterias/aislamiento & purificación , Cianobacterias/metabolismo , Genoma Bacteriano , Lagos/microbiología , Fenotipo , Filogenia , Plancton/genética , Plancton/aislamiento & purificaciónRESUMEN
BACKGROUND: Routine culture-based diagnosis of Pseudomonas aeruginosa lung infection in Cystic Fibrosis (CF) patients can be hampered by the phenotypic variability of the microorganism, including its transition to a Viable But Non-Culturable (VBNC) state. The aim of this study was to validate an ecfX-targeting qPCR protocol developed to detect all viable P. aeruginosa bacteria and to identify VBNC forms in CF sputum samples. METHODS: The study involved 115 P. aeruginosa strains of different origins and 10 non-P. aeruginosa strains and 88 CF sputum samples, 41 Culture-Positive (CP) and 47 Culture-Negative (CN). Spiking assays were performed using scalar dilutions of a mixture of live and dead P. aeruginosa ATCC 9027 and a pooled P. aeruginosa-free sputum batch. Total DNA from sputum samples was extracted by a commercial kit, whereas a crude extract was obtained from the broth cultures. Extracellular DNA (eDNA) interference was evaluated by comparing the qPCR counts obtained from DNase-treated and untreated aliquots of the same samples. The statistical significance of the results was assessed by the Wilcoxon test and Student's t test. RESULTS: The newly-developed qPCR protocol identified 96.6% of the P. aeruginosa isolates; no amplification was obtained with strains belonging to different species. Spiking assays supported protocol reliability, since counts always matched the amount of live bacteria, thus excluding the interference of dead cells and eDNA. The protocol sensitivity threshold was 70 cells/ml of the original sample. Moreover, qPCR detected P. aeruginosa in 9/47 CN samples and showed higher bacterial counts compared with the culture method in 10/41 CP samples. CONCLUSIONS: Our findings demonstrate the reliability of the newly-developed qPCR protocol and further highlight the need for harnessing a non-culture approach to achieve an accurate microbiological diagnosis of P. aeruginosa CF lung infection and a greater understanding of its evolution.
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Fibrosis Quística/microbiología , Viabilidad Microbiana , Técnicas Microbiológicas , Infecciones por Pseudomonas/diagnóstico , Pseudomonas aeruginosa , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Fibrosis Quística/complicaciones , Femenino , Humanos , Límite de Detección , Pulmón/microbiología , Masculino , Técnicas Microbiológicas/normas , Infecciones por Pseudomonas/complicaciones , Infecciones por Pseudomonas/microbiología , Pseudomonas aeruginosa/citología , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/aislamiento & purificación , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Esputo/microbiologíaRESUMEN
Objectives: To analyse the recombination events associated with conjugal mobilization of two multiresistance plasmids, pRUM17i48 and pLAG (formerly named pDO1-like), from Enterococcus faecium 17i48 to Enterococcus faecalis JH2-2. Methods: The plasmids from two E. faecalis transconjugants (JH-4T, tetracycline resistant, and JH-8E, erythromycin resistant) and from the E. faecium donor (also carrying a pHTß-like conjugative plasmid, named pHTß17i48) were investigated by several methods, including PCR mapping and sequencing, S1-PFGE followed by Southern blotting and hybridization, and WGS. Results: Two locations of repApHTß were detected in both transconjugants, one on a â¼50 kb plasmid (as in the donor) and the other on plasmids of larger sizes. In JH-4T, WGS disclosed an 88.6 kb plasmid resulting from the recombination of pHTß17i48 (â¼50 kb) and a new plasmid, named pLAG (35.3 kb), carrying the tet(M), tet(L), lsa(E), lnu(B), spw and aadE resistance genes. In JH-8E, a 75 kb plasmid resulting from the recombination of pHTß17i48 and pRUM17i48 was observed. In both cases, the cointegrates were apparently derived from replicative transposition of an IS1216 present in each of the multiresistance plasmids into pHTß17i48. The cointegrates could resolve to yield the multiresistance plasmids and a pHTß17i48 derivative carrying an IS1216 (unlike the pHTß17i48 of the donor). Conclusions: Our results completed the characterization of the multiresistance plasmids carried by the E. faecium 17i48, confirming the role of pHT plasmids in the mobilization of non-conjugative antibiotic resistance elements among enterococci. Results also revealed that mobilization to E. faecalis was associated with the generation of cointegrate plasmids promoted by IS1216-mediated transposition.
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Antibacterianos/farmacología , Conjugación Genética , Farmacorresistencia Bacteriana Múltiple , Enterococcus faecalis/genética , Enterococcus faecium/genética , Plásmidos/efectos de los fármacos , Recombinación Genética , Southern Blotting , ADN Bacteriano/genética , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecium/efectos de los fármacos , Eritromicina/farmacología , Genes Bacterianos/efectos de los fármacos , Humanos , Reacción en Cadena de la Polimerasa , Resistencia a la Tetraciclina/genéticaRESUMEN
Planktonic cyanobacteria belonging to the genus Synechococcus are ubiquitously distributed in marine and fresh waters, substantially contributing to total carbon fixation on a global scale. While their ecological relevance is acknowledged, increasing resolution in molecular techniques allows disentangling cyanobacteria's role at the micro-scale, where complex microbial interactions may drive the overall community assembly. The interplay between phylogenetically different Synechococcus clades and their associated bacterial communities can affect their ecological fate and susceptibility to protistan predation. In this study, we experimentally promoted different levels of ecological interaction by mixing two Synechococcus ribotypes (MW101C3 and LL) and their associated bacteria, with and without a nanoflagellate grazer (Poterioochromonas sp.) in laboratory cultures. The beta-diversity of the Synechococcus-associated microbiome in laboratory cultures indicated that the presence of the LL ribotype was the main factor determining community composition changes (41% of total variance), and prevailed over the effect of protistan predation (18% of total variance). Our outcomes also showed that species coexistence and predation may promote microbial diversity, thus highlighting the underrated ecological relevance of such micro-scale factors.
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Rasgos de la Historia de Vida , Microbiota , Synechococcus/fisiología , Chrysophyta/fisiología , Cadena Alimentaria , Ribotipificación , Synechococcus/genéticaRESUMEN
Antibiotic resistance genes (ARGs) are increasingly appreciated to be important as micropollutants. Indirectly produced by human activities, they are released into the environment, as they are untargeted by conventional wastewater treatments. In order to understand the fate of ARGs and of other resistant forms (e.g., phenotypical adaptations) in urban wastewater treatment plants (WWTPs), we monitored three WWTPs with different disinfection processes (chlorine, peracetic acid (PAA), and ultraviolet light (UV)). We monitored WWTPs influx and pre- and postdisinfection effluent over 24 h, followed by incubation experiments lasting for 96 h. We measured bacterial abundance, size distribution and aggregational behavior, the proportion of intact (active) cells, and the abundances of four ARGs and of the mobile element integron1. While all the predisinfection treatments of all WWTPs removed the majority of bacteria and of associated ARGs, of the disinfection processes only PAA efficiently removed bacterial cells. However, the stress imposed by PAA selected for bacterial aggregates and, similarly to chlorine, stimulated the selection of ARGs during the incubation experiment. This suggests disinfections based on chemically aggressive destruction of bacterial cell structures can promote a residual microbial community that is more resistant to antibiotics and, given the altered aggregational behavior, to competitive stress in nature.
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Desinfección , Aguas Residuales/química , Antibacterianos/farmacología , Bacterias/genética , Farmacorresistencia Microbiana/genética , Genes Bacterianos/efectos de los fármacos , HumanosRESUMEN
The fate of antibiotic resistance genes (ARGs) in environmental microbial communities is of primary concern as prodromal of a potential transfer to pathogenic bacteria. Although of diverse origin, the persistence of ARGs in aquatic environments is highly influenced by anthropic activities, allowing potential control actions in well-studied environments. However, knowledge of abundance and space-time distribution of ARGs in ecosystems is still scarce. Using quantitative real-time PCR, we investigated the presence and the abundance of twelve ARGs (against tetracyclines, ß-lactams, aminoglycosides, quinolones and sulphonamides) at different sampling sites, depths and seasons, in Lake Maggiore, a large subalpine lake, and in the area of its watershed. We then evaluated the correlation between each ARG and a number of ecological parameters in the water column in the deepest part of the lake. Our results suggest the constitutive presence of at least four ARGs within the bacterial community with a high proportion of bacteria potentially resistant to tetracyclines and sulphonamides. The presence of these ARGs was independent of the total bacterial density and temperature. The dynamics of tet(A) and sulII genes were, however, positively correlated with dissolved oxygen and negatively to chlorophyll a, suggesting that the resistant microbes inhabit specific niches. These observations indicate that the lake is a reservoir of antibiotic resistances, highlighting the need of a deeper understanding of the sources of ARGs and the factors allowing their persistence in waters.
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Bacterias/genética , Farmacorresistencia Bacteriana/genética , Genes Bacterianos , Lagos/microbiología , ADN Bacteriano/genética , Italia , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ADN , SuizaRESUMEN
Aeromonas hydrophila is an aquatic bacterium responsible for several human illnesses. The aim of this work was to investigate the survival ability and virulence expression of two strains from different sources (fish, strain 87 and surface water, strain LS) maintained in a seawater microcosm. The strains were analyzed for the total and viable bacterial counts, adhesion ability to Hep-2 cells and aerA gene expression by qPCR throughout the experiment (35 days). Both strains reached a putative VBNC state and lost adhesive properties but exhibited a different behavior in the expression of aerA. This could be due to the different origin of the two strains; the former adapted to a habitat rich of nutrient and the latter already used to survive in a more hostile environment. Moreover, our results indicate that the quantitative determination of aerA mRNA can be a useful indicator of virulence expression under stress conditions.
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Aeromonas hydrophila/fisiología , Aeromonas hydrophila/patogenicidad , Regulación Bacteriana de la Expresión Génica/fisiología , Viabilidad Microbiana , Factores de Virulencia/genética , Aeromonas hydrophila/genética , Animales , Carga Bacteriana , Toxinas Bacterianas/genética , Adhesión Celular/fisiología , Peces/microbiología , Células Hep G2 , Humanos , Proteínas Citotóxicas Formadoras de Poros/genética , Agua de Mar/microbiologíaRESUMEN
The dual nature of Legionella pneumophila enables its survival in free and intracellular environments and underpins its infection and spread mechanisms. Experiments using bacterial cultures and improved RTqPCR protocols were devised to gain fresh insights into the role of biofilm in protecting the replicative form of L. pneumophila. mip gene expression was used as a marker of virulence in sessile (biofilm-bound) and planktonic (free-floating) cells of L. pneumophila serotype 1 ATCC 33152. The ratio of mip gene expression to transcriptionally active Legionella cells increased both in sessile and free-floating cells demonstrating an up-regulation of mip gene under nutrient depletion. However, a different trend was observed between the two forms, in planktonic cells the mip gene expression/transcriptionally active Legionella cells increased until the end of the experiment, while in the biofilm such increase was observed at the end of the experiment. These findings suggest a possible association between the switch to the transmissive phase of Legionella and a mip up-regulation and a role for biofilm in preserving Legionella cells in replicative form. Moreover, it has been shown that improved RTqPCR protocols are valuable tools to explore bacterial virulence.
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Biopelículas/crecimiento & desarrollo , Legionella pneumophila/fisiología , Viabilidad Microbiana , Proteínas Bacterianas/análisis , Proteínas Bacterianas/genética , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica , Legionella pneumophila/genética , Legionella pneumophila/crecimiento & desarrollo , Isomerasa de Peptidilprolil/análisis , Isomerasa de Peptidilprolil/genética , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The presence of antimicrobial resistance genes (ARGs) in the microbiome of freshwater communities is a consequence of thousands of years of evolution but also of the pressure exerted by anthropogenic activities, with potential negative impact on environmental and human health. In this study, we investigated the distribution of ARGs in Lake Tanganyika (LT)'s water column to define the resistome of this ancient lake. Additionally, we compared the resistome of LT with that of Lake Baikal (LB), the oldest known lake with different environmental characteristics and a lower anthropogenic pollution than LT. We found that richness and abundance of several antimicrobial resistance classes were higher in the deep water layers in both lakes. LT Kigoma region, known for its higher anthropogenic pollution, showed a greater richness and number of ARG positive MAGs compared to Mahale. Our results provide a comprehensive understanding of the antimicrobial resistome of LT and underscore its importance as reservoir of antimicrobial resistance. In particular, the deepest water layers of LT are the main repository of diverse ARGs, mirroring what was observed in LB and in other aquatic ecosystems. These findings suggest that the deep waters might play a crucial role in the preservation of ARGs in aquatic ecosystems.
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Antiinfecciosos , Microbiota , Humanos , Lagos , Agua , Tanzanía , Genes Bacterianos , AntibacterianosRESUMEN
The escalating presence of antibiotic-resistant bacteria (ARB) in aquatic ecosystems underscores the critical role of wastewater treatment plants (WWTPs) in mitigating antibiotic resistance. Disinfection is the final, pivotal step in WWTPs, and it is essential to control the dissemination of ARB before water discharge. This study utilized both phenotypic analysis and transcriptome (RNA-seq) approach to investigate the efficiency and mechanisms of disinfection using chlorination, ultraviolet (UV), and peracetic acid (PAA) on multidrug-resistant bacteria (MRB). Our results demonstrated that the use of 100 mg min L-1 of chlorine, 8.19 mJ cm-2 of UV irradiation or 50 min mg L-1 of PAA significantly reduced the abundance of MRB. Intriguingly, RNA-seq clarified distinct mechanisms of chlorination and UV disinfection. UV radiation triggered the SOS response to cope with DNA damage, induced the expression of multi-drug resistance genes by increasing the expression of efflux pump transporters. UV radiation also promoted the absorption of iron through chelation and transportation to participate in various cell life processes. Chlorination, on the other hand, significantly up-regulated osmotic response elements, including the synthesis of glycine betaine, iron-sulfur clusters, and related transporters. Both chlorination and UV significantly down-regulated key metabolic pathways (P < 0.05), inhibiting the process of amino acid synthesis and energy metabolism. Imbalance in energy homeostasis was the most important factor leading to cytotoxicity. These results provide useful insights into optimizing the wastewater disinfection process in order to prevent the dissemination of ARB in aquatic environment.
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Aguas Residuales , Purificación del Agua , Desinfección/métodos , Ecosistema , Antagonistas de Receptores de Angiotensina , Inhibidores de la Enzima Convertidora de Angiotensina , Ácido Peracético , Rayos Ultravioleta , Bacterias , Hierro , Purificación del Agua/métodos , Antibacterianos/farmacologíaRESUMEN
Microplastic particles are persistent micropollutants that provide a substrate for the growth of bacterial biofilms, posing a threat to the environment. This study explores the changes in commercially available food containers made of conventional (polypropylene PP, polyethylene terephthalate PET), innovative biodegradable (Mater-Bi) and natural (wood and cellulose) materials, when introduced in the surface waters of Lake Maggiore for 43 days. Spectral changes revealed by FT-IR spectroscopy in PET and Mater-Bi, and changes in thermal properties of all human-made material tested indicated a degradation process occurred during environmental exposure. Despite similar bacterial richness, biofilms on PET, PP, and Mater-Bi differed from natural material biofilms and the planktonic community. Human-made material communities showed a higher proportion of potential pathogens, with PET and PP also exhibiting increased abundances of antibiotic resistance genes. Overall, these findings stress the need for dedicated strategies to curb the spread of human-made polymers in freshwaters, including innovative materials that, due to their biodegradable properties, might be perceived less hazardous for the environment.
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Plásticos Biodegradables , Humanos , Plásticos , Plancton , Espectroscopía Infrarroja por Transformada de Fourier , Polipropilenos , Microplásticos , BacteriasRESUMEN
Wastewater treatment plants are hotspots for the release of antimicrobial resistant pathogenic bacteria into aquatic ecosystems, significantly contributing to the cycle of antimicrobial resistance. Special attention should be paid to antimicrobial resistant ESKAPE bacteria, which have been identified as high-priority targets for control measures. Among them, Klebsiella pneumoniae is particularly noteworthy. In this study, we collected wastewater samples from the inlet, sedimentation tank, and effluent water of a wastewater treatment plant in June, July, October, and November of 2018. We detected and characterized 42 K. pneumoniae strains using whole genome sequencing (15 from the inlet, 8 from the sedimentation tank, and 19 from the effluent). Additionally, the strains were tested for their antimicrobial resistance phenotype. Using whole genome sequencing no distinct patterns were observed in terms of their genetic profiles. All strains were resistant to tetracycline, meanwhile 60%, 47%, and 37.5% of strains isolated from the inlet, sedimentation tank, and effluent, respectively, were multidrug resistant. Some of the multidrug resistant isolates were also resistant to colistin, and nearly all tested positive for the eptB and arnT genes, which are associated with polymyxin resistance. Various antimicrobial resistance genes were linked to mobile genetic elements, and they did not correlate with detected virulence groups or defense systems. Overall, our results, although not quantitative, highlight that multidrug resistant K. pneumoniae strains, including those resistant to colistin and genetically unrelated, being discharged into aquatic ecosystems from wastewater treatment plants. This suggests the necessity of monitoring aimed at genetically characterizing these pathogenic bacteria.
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Introduction: Constructed wetlands (CWs) are nature-based solutions for wastewater treatment where the root system microbiome plays a key role in terms of nutrient and pollutant removal. Nonetheless, little is known on plant-microbe interactions and bacterial population selection in CWs, which are mostly characterized in terms of engineering aspects. Methods: Here, cultivation-independent and cultivation-based analyses were applied to study the bacterial communities associated to the root systems of Phragmites australis and Typha domingensis co-occurring in the same cell of a CW receiving primary treated wastewaters. Results and discussion: Two endophytic bacteria collections (n = 156) were established aiming to find novel strains for microbial-assisted phytodepuration, however basing on their taxonomy the possible use of these strains was limited by their low degrading potential and/or for risks related to the One-Health concept. A sharp differentiation arose between the P. australis and T. domingensis collections, mainly represented by lactic acid bacteria (98%) and Enterobacteriaceae (69%), respectively. Hence, 16S rRNA amplicon sequencing was used to disentangle the microbiome composition in the root system fractions collected at increasing distance from the root surface. Both the fraction type and the plant species were recognized as drivers of the bacterial community structure. Moreover, differential abundance analysis revealed that, in all fractions, several bacteria families were significantly and differentially enriched in P. australis or in T. domingensis. CWs have been also reported as interesting options for the removal of emerging contaminants (e.g, antibiotic resistance genes, ARGs). In this study, ARGs were mostly present in the rhizosphere of both plant species, compared to the other analyzed fractions. Notably, qPCR data showed that ARGs (i.e., ermB, bla TEM, tetA) and intl1 gene (integrase gene of the class 1 integrons) were significantly higher in Phragmites than Typha rhizospheres, suggesting that macrophyte species growing in CWs can display a different ability to remove ARGs from wastewater. Overall, the results suggest the importance to consider the plant-microbiome interactions, besides engineering aspects, to select the most suitable species when designing phytodepuration systems.
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Antibiotic resistance genes (ARGs) are abundant in aquatic ecosystems affected by human activities. Understanding the fate of ARGs across different ecosystems is essential because of the significant role aquatic environments play in the cycle of antibiotic resistance. We quantified selected ARGs in Lake Maggiore, its main tributaries, and the effluent of the main wastewater treatment plant (WWTP) discharging directly into the lake. We linked their dynamics to the different anthropogenic impacts in each tributary's watershed. The dynamics of tetA in the lake were influenced by those of the rivers and the WWTP effluent, and by the concentration of N-NH4, related to anthropogenic pollution, while sul2 abundance in the lake was not influenced by any water inflow. The dynamics of the different ARGs varied across the different rivers. Rivers with watersheds characterized by high population density, touristic activities, and secondary industries released more ARGs, while ermB correlated with higher numbers of primary industries. This study suggests a limited contribution of treated wastewater in the spread of ARGs, indicating as prevalent origin other sources of pollution, calling for a reconsideration on what are considered the major sources of ARGs into the environment.
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Antibacterianos , Genes Bacterianos , Humanos , Antibacterianos/farmacología , Antibacterianos/análisis , Lagos , Ecosistema , Farmacorresistencia Microbiana/genética , RíosRESUMEN
Petroleum-based microplastic particles (MPs) are carriers of antimicrobial resistance genes (ARGs) in aquatic environments, influencing the selection and spread of antimicrobial resistance. This research characterized MP and natural organic particle (NOP) bacterial communities and resistomes in the Tyrrhenian Sea, a region impacted by plastic pollution and climate change. MP and NOP bacterial communities were similar but different from the free-living planktonic communities. Likewise, MP and NOP ARG abundances were similar but different (higher) from the planktonic communities. MP and NOP metagenome-assembled genomes contained ARGs associated with mobile genetic elements and exhibited co-occurrence with metal resistance genes. Overall, these findings show that MPs and NOPs harbor potential pathogenic and antimicrobial resistant bacteria, which can aid in the spread of antimicrobial resistance. Further, petroleum-based MPs do not represent novel ecological niches for allochthonous bacteria; rather, they synergize with NOPs, collectively facilitating the spread of antimicrobial resistance in marine ecosystems.
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Bacterias , Microplásticos , Bacterias/genética , Bacterias/efectos de los fármacos , Microplásticos/toxicidad , Contaminantes Químicos del Agua/análisis , Microbiota/efectos de los fármacos , Farmacorresistencia Bacteriana/genética , Monitoreo del Ambiente , Agua de Mar/microbiología , Agua de Mar/químicaRESUMEN
Aquatic ecosystems are crucial in the antimicrobial resistance cycle. While intracellular DNA has been extensively studied to understand human activity's impact on antimicrobial resistance gene (ARG) dissemination, extracellular DNA is frequently overlooked. This study examines the effect of anthropogenic water pollution on microbial community diversity, the resistome, and ARG dissemination. We analyzed intracellular and extracellular DNA from wastewater treatment plant effluents and lake surface water by shotgun sequencing. We also conducted experiments to evaluate anthropogenic pollution's effect on transforming extracellular DNA (using Gfp-plasmids carrying ARGs) within a natural microbial community. Chemical analysis showed treated wastewater had higher anthropogenic pollution-related parameters than lake water. The richness of microbial community, antimicrobial resistome, and high-risk ARGs was greater in treated wastewaters than in lake waters both for intracellular and extracellular DNA. Except for the high-risk ARGs, richness was significantly higher in intracellular than in extracellular DNA. Several ARGs were associated with mobile genetic elements and located on plasmids. Furthermore, Gfp-plasmid transformation within a natural microbial community was enhanced by anthropogenic pollution levels. Our findings underscore anthropogenic pollution's pivotal role in shaping microbial communities and their antimicrobial resistome. Additionally, it may facilitate ARG dissemination through extracellular DNA plasmid uptake.