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In recent years, microbiomes and their potential applications for human, animal or plant health, food production and environmental management came into the spotlight of major national and international policies and strategies. This has been accompanied by substantial R&D investments in both public and private sectors, with an increasing number of products entering the market. Despite widespread agreement on the potential of microbiomes and their uses across disciplines, stakeholders and countries, there is no consensus on what defines a microbiome application. This often results in non-comprehensive communication or insufficient documentation making commercialisation and acceptance of the novel products challenging. To showcase the complexity of this issue we discuss two selected, well-established applications and propose criteria defining a microbiome application and their conditions of use for clear communication, facilitating suitable regulatory frameworks and building trust among stakeholders.
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Microbiota , Humanos , AnimalesRESUMEN
Wastewater surveillance of SARS-CoV-2 has emerged as a critical tool for tracking the spread of COVID-19. In addition to estimating the relative case numbers using quantitative PCR, SARS-CoV-2 genomic RNA can be extracted from wastewater and sequenced. There are many existing techniques for using the sequenced RNA to determine the relative abundance of known lineages in a sample. However, it is very challenging to predict novel lineages from wastewater data due to its mixed composition and unreliable genomic coverage. In this work, we present a novel technique based on non-negative matrix factorization which is able to reconstruct lineage definitions by analyzing data from across different samples. We test the method both on synthetic and real wastewater sequencing data. We show that the technique is able to determine major lineages such as Omicron and Delta as well as sub-lineages such as BA.5.2.1. We provide a method for determining emerging lineages in wastewater without the need for genomic data from clinical samples. This could be used for routine monitoring of SARS-CoV-2 as well as other emerging viral pathogens in wastewater. Additionally, it may be used to determine more full-genome sequences for viruses with fewer available genomes.
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COVID-19 , Genoma Viral , Filogenia , ARN Viral , SARS-CoV-2 , Aguas Residuales , Aguas Residuales/virología , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , COVID-19/virología , Humanos , ARN Viral/genéticaRESUMEN
During the COVID-19 pandemic, the Province of Ontario, Canada, launched a wastewater surveillance program to monitor SARS-CoV-2, inspired by the early work and successful forecasts of COVID-19 waves in the city of Ottawa, Ontario. This manuscript presents a dataset from January 1, 2021, to March 31, 2023, with RT-qPCR results for SARS-CoV-2 genes and PMMoV from 107 sites across all 34 public health units in Ontario, covering 72% of the province's and 26.2% of Canada's population. Sampling occurred 2-7 times weekly, including geographical coordinates, serviced populations, physico-chemical water characteristics, and flowrates. In doing so, this manuscript ensures data availability and metadata preservation to support future research and epidemic preparedness through detailed analyses and modeling. The dataset has been crucial for public health in tracking disease locally, especially with the rise of the Omicron variant and the decline in clinical testing, highlighting wastewater-based surveillance's role in estimating disease incidence in Ontario.
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COVID-19 , SARS-CoV-2 , Aguas Residuales , Ontario/epidemiología , COVID-19/epidemiología , Aguas Residuales/virología , Humanos , Pandemias , Carga ViralRESUMEN
Wastewater-based surveillance (WBS) is an important epidemiological and public health tool for tracking pathogens across the scale of a building, neighbourhood, city, or region. WBS gained widespread adoption globally during the SARS-CoV-2 pandemic for estimating community infection levels by qPCR. Sequencing pathogen genes or genomes from wastewater adds information about pathogen genetic diversity, which can be used to identify viral lineages (including variants of concern) that are circulating in a local population. Capturing the genetic diversity by WBS sequencing is not trivial, as wastewater samples often contain a diverse mixture of viral lineages with real mutations and sequencing errors, which must be deconvoluted computationally from short sequencing reads. In this study we assess nine different computational tools that have recently been developed to address this challenge. We simulated 100 wastewater sequence samples consisting of SARS-CoV-2 BA.1, BA.2, and Delta lineages, in various mixtures, as well as a Delta-Omicron recombinant and a synthetic 'novel' lineage. Most tools performed well in identifying the true lineages present and estimating their relative abundances and were generally robust to variation in sequencing depth and read length. While many tools identified lineages present down to 1â% frequency, results were more reliable above a 5â% threshold. The presence of an unknown synthetic lineage, which represents an unclassified SARS-CoV-2 lineage, increases the error in relative abundance estimates of other lineages, but the magnitude of this effect was small for most tools. The tools also varied in how they labelled novel synthetic lineages and recombinants. While our simulated dataset represents just one of many possible use cases for these methods, we hope it helps users understand potential sources of error or bias in wastewater sequencing analysis and to appreciate the commonalities and differences across methods.
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COVID-19 , Genoma Viral , SARS-CoV-2 , Aguas Residuales , Aguas Residuales/virología , SARS-CoV-2/genética , SARS-CoV-2/clasificación , COVID-19/virología , COVID-19/epidemiología , Humanos , Biología Computacional/métodos , Genómica/métodos , Monitoreo Epidemiológico Basado en Aguas Residuales , FilogeniaRESUMEN
Tomato Brown Rugose Fruit Virus (ToBRFV) is a plant pathogen that infects important Solanaceae crop species and can dramatically reduce tomato crop yields. The ToBRFV has rapidly spread around the globe due to its ability to escape detection by antiviral host genes which confer resistance to other tobamoviruses in tomato plants. The development of robust and reproducible methods for detecting viruses in the environment aids in the tracking and reduction of pathogen transmission. We detected ToBRFV in municipal wastewater influent (WWI) samples, likely due to its presence in human waste, demonstrating a widespread distribution of ToBRFV in WWI throughout Ontario, Canada. To aid in global ToBRFV surveillance efforts, we developed a tiled amplicon approach to sequence and track the evolution of ToBRFV genomes in municipal WWI. Our assay recovers 95.7% of the 6393 bp ToBRFV RefSeq genome, omitting the terminal 5' and 3' ends. We demonstrate that our sequencing assay is a robust, sensitive, and highly specific method for recovering ToBRFV genomes. Our ToBRFV assay was developed using existing ARTIC Network resources, including primer design, sequencing library prep, and read analysis. Additionally, we adapted our lineage abundance estimation tool, Alcov, to estimate the abundance of ToBRFV clades in samples.
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Solanum lycopersicum , Tobamovirus , Purificación del Agua , Humanos , Ontario , Frutas , Tobamovirus/genéticaRESUMEN
Wastewater surveillance of coronavirus disease 2019 (COVID-19) commonly applies reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to quantify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentrations in wastewater over time. In most applications worldwide, maximal sensitivity and specificity of RT-qPCR has been achieved, in part, by monitoring two or more genomic loci of SARS-CoV-2. In Ontario, Canada, the provincial Wastewater Surveillance Initiative reports the average copies of the CDC N1 and N2 loci normalized to the fecal biomarker pepper mild mottle virus. In November 2021, the emergence of the Omicron variant of concern, harboring a C28311T mutation within the CDC N1 probe region, challenged the accuracy of the consensus between the RT-qPCR measurements of the N1 and N2 loci of SARS-CoV-2. In this study, we developed and applied a novel real-time dual loci quality assurance and control framework based on the relative difference between the loci measurements to the City of Ottawa dataset to identify a loss of sensitivity of the N1 assay in the period from July 10, 2022 to January 31, 2023. Further analysis via sequencing and allele-specific RT-qPCR revealed a high proportion of mutations C28312T and A28330G during the study period, both in the City of Ottawa and across the province. It is hypothesized that nucleotide mutations in the probe region, especially A28330G, led to inefficient annealing, resulting in reduction in sensitivity and accuracy of the N1 assay. This study highlights the importance of implementing quality assurance and control criteria to continually evaluate, in near real-time, the accuracy of the signal produced in wastewater surveillance applications that rely on detection of pathogens whose genomes undergo high rates of mutation.
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Monitoreo Epidemiológico Basado en Aguas Residuales , Aguas Residuales , Alelos , Mutación , Ontario/epidemiología , SARS-CoV-2/genética , ARN Viral/genéticaRESUMEN
Microbiomes have highly important roles for ecosystem functioning and carry out key functions that support planetary health, including nutrient cycling, climate regulation, and water filtration. Microbiomes are also intimately associated with complex multicellular organisms such as humans, other animals, plants, and insects and perform crucial roles for the health of their hosts. Although we are starting to understand that microbiomes in different systems are interconnected, there is still a poor understanding of microbiome transfer and connectivity. In this review we show how microbiomes are connected within and transferred between different habitats and discuss the functional consequences of these connections. Microbiome transfer occurs between and within abiotic (e.g., air, soil, and water) and biotic environments, and can either be mediated through different vectors (e.g., insects or food) or direct interactions. Such transfer processes may also include the transmission of pathogens or antibiotic resistance genes. However, here, we highlight the fact that microbiome transmission can have positive effects on planetary and human health, where transmitted microorganisms potentially providing novel functions may be important for the adaptation of ecosystems.
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Microbiota , Planetas , Animales , Humanos , Microbiología del Suelo , Microbiota/fisiología , Suelo , AguaRESUMEN
Production of polyhydroxyalkanoates (PHAs) with high concentration of carboxylate, that was accumulated from solid state fermentation (SSF) of food waste (FW), was tested using Pseudomonas putida strain KT2440. Mixed-culture SSF of FW supplied in a high concentration of carboxylate, which caused a high PHA production of 0.56 g PHA/g CDM under nutrients control. Interestingly, this high PHA fraction in CDM was almost constant at 0.55 g PHA/g CDM even under high nutrients concentration (25 mM NH4+), probably due to high reducing power maintained by high carboxylate concentration. PHA characterization indicated that the dominant PHA building block produced was 3-hydroxybutyrate, followed by 3-hydroxy-2-methylvalerate and 3-hydroxyhenxanoate. Carboxylate profiles before and after PHA production suggested that acetate, butyrate, and propionate were the main precursors to PHA via several metabolic pathways. Our result support that mixed culture SSF of FW for high concentration carboxylate and P. putida for PHA production enables sustainable production of PHA in cost-effective manners.
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Polihidroxialcanoatos , Pseudomonas putida , Eliminación de Residuos , Pseudomonas putida/metabolismo , Polihidroxialcanoatos/metabolismo , Alimentos , Fermentación , Ácidos CarboxílicosRESUMEN
Research on the role of gut microbiota in behavior has grown dramatically. The probiotic L. reuteri can alter social and stress-related behaviors - yet, the underlying mechanisms remain largely unknown. Although traditional laboratory rodents provide a foundation for examining the role of L. reuteri on the gut-brain axis, they do not naturally display a wide variety of social behaviors. Using the highly-social, monogamous prairie vole (Microtus ochrogaster), we examined the effects of L. reuteri administration on behaviors, neurochemical marker expression, and gut-microbiome composition. Females, but not males, treated with live L. reuteri displayed lower levels of social affiliation compared to those treated with heat-killed L. reuteri. Overall, females displayed a lower level of anxiety-like behaviors than males. Live L. reuteri-treated females had lower expression of corticotrophin releasing factor (CRF) and CRF type-2-receptor in the nucleus accumbens, and lower vasopressin 1a-receptor in the paraventricular nucleus of the hypothalamus (PVN), but increased CRF in the PVN. There were both baseline sex differences and sex-by-treatment differences in gut microbiome composition. Live L. reuteri increased the abundance of several taxa, including Enterobacteriaceae, Lachnospiraceae NK4A136, and Treponema. Interestingly, heat-killed L. reuteri increased abundance of the beneficial taxa Bifidobacteriaceae and Blautia. There were significant correlations between changes in microbiota, brain neurochemical markers, and behaviors. Our data indicate that L. reuteri impacts gut microbiota, gut-brain axis and behaviors in a sex-specific manner in socially-monogamous prairie voles. This demonstrates the utility of the prairie vole model for further examining causal impacts of microbiome on brain and behavior.
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The ability to produce high-value products using bacteria will increasingly rely on continued research to make large-scale bacterial fermentation cost-efficient. Engineering bacteria to use alternate carbon sources as feedstock provides an opportunity to reduce production costs. Using inexpensive carbon sources from various forms of waste provides an opportunity to substantially reduce feedstock costs. Functional carbon metabolism pathways can be identified by the introduction of metagenomic libraries into the organism of interest followed by screening for the desired phenotype. We present here a method to transfer metagenomic libraries from E. coli to Pseudomonas alloputida, followed by screening for use of galactose as a sole carbon source.
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Carbono , Microbiota , Carbono/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Metagenoma , Metagenómica , FermentaciónRESUMEN
The resilience and low cost of plastics has made their usage ubiquitous, but is also the cause of their prevalence and longevity as waste. Plastic pollution has become a great concern to the health and wellbeing of ecosystems around the world; microplastics are a particular threat, due to their high mobility, ease of ingestion by wildlife, and ability to adsorb and carry toxic contaminants. Material flow analysis has been widely applied to examine stocks and flows of materials in other industries, and has more recently been applied to plastics to examine areas where waste can reach the environment. However, while much research has gone into the environmental fate of microplastics, degradation strategies have been a lesser focus, and material flow analysis of microplastics has suffered from lack of data. Furthermore, the variety of plastics, their additives, and any contaminants pose a significant challenge in degrading (and not merely fragmenting) microplastic particles. This review discusses the current degradation strategies and solutions for dealing with existing and newly-generated microplastic waste along with examining the status of microplastics-based material flow analysis, which are critical for evaluating the possibility of incorporating microplastic waste into a circular economy. The degradation strategies are critically examined, identifying challenges and current trends, as well as important considerations that are frequently under-reported. An emphasis is placed on identifying missing data or information in both material flow analysis and degradation methods that could prove crucial in improving understanding of microplastic flows, as well as optimizing degradation strategies and minimizing any negative environmental impact.
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Microplásticos , Contaminantes Químicos del Agua , Plásticos/toxicidad , Ecosistema , Contaminantes Químicos del Agua/análisis , Contaminación Ambiental , Monitoreo del AmbienteRESUMEN
Bacterial cells are widely used to produce value-added products due to their versatility, ease of manipulation, and the abundance of genome engineering tools. However, the efficiency of producing these desired biomolecules is often hindered by the cells' own metabolism, genetic instability, and the toxicity of the product. To overcome these challenges, genome reductions have been performed, making strains with the potential of serving as chassis for downstream applications. Here we review the current technologies that enable the design and construction of such reduced-genome bacteria as well as the challenges that limit their assembly and applicability. While genomic reductions have shown improvement of many cellular characteristics, a major challenge still exists in constructing these cells efficiently and rapidly. Computational tools have been created in attempts at minimizing the time needed to design these organisms, but gaps still exist in modelling these reductions in silico. Genomic reductions are a promising avenue for improving the production of value-added products, constructing chassis cells, and for uncovering cellular function but are currently limited by their time-consuming construction methods. With improvements to and the creation of novel genome editing tools and in silico models, these approaches could be combined to expedite this process and create more streamlined and efficient cell factories.
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The overarching biological impact of microbiomes on their hosts, and more generally their environment, reflects the co-evolution of a mutualistic symbiosis, generating fitness for both. Knowledge of microbiomes, their systemic role, interactions, and impact grows exponentially. When a research field of importance for planetary health evolves so rapidly, it is essential to consider it from an ethical holistic perspective. However, to date, the topic of microbiome ethics has received relatively little attention considering its importance. Here, ethical analysis of microbiome research, innovation, use, and potential impact is structured around the four cornerstone principles of ethics: Do Good; Don't Harm; Respect; Act Justly. This simple, but not simplistic approach allows ethical issues to be communicative and operational. The essence of the paper is captured in a set of eleven microbiome ethics recommendations, e.g., proposing gut microbiome status as common global heritage, similar to the internationally agreed status of major food crops.
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Nanochitin whisker (NC) is an advanced nanobiomaterial with novel physicochemical and biological properties. Fusarium pseudograminearum (Fpg) is an important pathogenic fungus causing wheat crown rot disease. This study explored the mode of action of NC against Fpg as a target microorganism. The effects of different treatments and concentrations of NC on the fungal growth and conidial germination were investigated by in vitro bioassay. The impacts of NC on cell structure integrity, membrane permeability, pathogenesis related key enzymes activity, and the mycotoxin production were examined by electron microscopy, fluorescence spectroscopy, IR spectroscopy, conductometry, and spectrophotometry, respectively. The results showed that NC significantly reduced hyphal growth, and the spore germination rate of Fpg declined by 33.0 % and 23.2 % when Fpg was treated with 30 and 300 µg/mL of NC, respectively. NC vigorously influenced structural stability of cell wall by destroying dextran structure, and strongly stimulated ergosterol production altering membrane integrity of the fungus. It reduced the activities of enzymes related to energy-supply like nicotinamide adenine dinucleotide oxidase and succinate dehydrogenase remarkably. The production of fungal mycotoxin deoxynivalenol was also decreased by NC. These findings provide an important basis for fully understanding the mechanism of nanobiomaterial in plant fungal disease control.
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Fusarium , Micotoxinas , Animales , Micotoxinas/farmacología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , VibrisasRESUMEN
Promoter engineering has been employed as a strategy to enhance and optimize the production of bio-products. Availability of promoters with predictable activities is needed for downstream application. However, whether promoter activity remains the same in different gene contexts remains unknown. Six consecutive promoters that have previously been determined to have different activity levels were used to construct six different versions of plasmid backbone pTH1227, followed by inserted genes encoding two polymer-producing enzymes. In some cases, promoter activity in the presence of inserted genes did not correspond to the reported activity levels in a previous study. After removing the inserted genes, the activity of these promoters returned to their previously reported level. These changes were further confirmed to occur at the transcriptional level. Polymer production using our newly constructed plasmids showed polymer accumulation levels corresponding to the promoter activity reported in our study. Our study demonstrated the importance of re-assessing promoter activity levels with regard to gene context, which could influence promoter activity, leading to different outcomes in downstream applications.
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Plásmidos , Plásmidos/genética , Regiones Promotoras GenéticasRESUMEN
The natural variation of multiple abiotic stresses in hyper-seasonal edaphic savanna provides a unique opportunity to study the rhizobacteriome community structure of plants adapted to climate change-like conditions in the humid tropics. In this study, we evaluated changes in soil, plant and rhizobacteriome community structure parameters across seasons (wet and dry) in two edaphic savannas (SV-1 and SV-5) using four dominant plant species. We then examined relationships between rhizobacteriome community structure and soil properties, plant biomass, and conventional and novel root traits. We further hypothesized that plants adapted to the Aripo Savanna had a core rhizobacteriome, which was specific to plant species and related to root foraging traits. Our results showed that cation exchange capacity (CEC) and the concentration of micronutrients (Fe, Cu and B) were the only soil factors that differed across savanna and season, respectively. Plant biomass traits were generally higher in the dry season, with a higher allocation to root growth in SV-5. Root traits were more plastic in SV-5, and network length-distribution was the only root trait which showed a consistent pattern of lower values in the dry season for three of the dominant plant species. Rhizobacterial community compositions were dominated by Proteobacteria and Acidobacteria, as well as WPS-2, which is dominant in extreme environments. We identified a shared core rhizobacteriome across plant species and savannas. Cation exchange capacity was a major driver of rhizobacterial community assemblies across savannas. Savanna-specific drivers of rhizobacterial community assemblies included CEC and Fe for SV-1, and CEC, TDS, NH4+, NO3-, Mn, K, and network length-distribution for SV-5. Plant factors on the microbiome were minimal, and host selectivity was mediated by the seasonal changes. We conclude that edaphoclimatic factors (soil and season) are the key determinants influencing rhizobacteriome community structure in multiple stressed-environments, which are ecologically similar to the Aripo Savanna.
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Ecosistema , Pradera , Biomasa , Plantas , SueloRESUMEN
Digital sequence information (DSI) has no clear definition. Numerous countries define DSI as a nonphysical genetic resources, such as genetic sequence data. Restricting free sharing of DSI is at odds with fundamental science core principles in disciplines like microbiology and molecular genetics. It has the potential to adversely affect international research collaborations.
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Difusión de la Información , Cooperación Internacional , Datos de Secuencia Molecular , IncertidumbreRESUMEN
Despite the risks that hydrocarbon contamination from pipeline leaks or train derailments impose on the health of peatlands in hydrocarbon production areas and transportation corridors, assessing the effect of such contaminations on the health and sustainability of peatlands has received little attention. This study investigates the impacts of hydrocarbons on peat microbial communities. Column experiments were conducted on non-aqueous phase liquid (NAPL) contaminated undisturbed peat core (0-35 cm) under static and fluctuating water table conditions. Water table fluctuations reduced residual NAPL saturation from 8.1-11.3% to 7.7-9.5%. Biodegradation of n-C8 and n-C12 along with oxidation of CH4 together produced high CO2 concentrations in the headspace. Clear patterns in dynamics in the microbial community structure were observed, with a more pronounced population growth. However, a significant loss of microbial richness was observed in contaminated columns. The result indicates that the phylum Proteobacteria benefited most from NAPL; however, their families differed between static and fluctuating water table conditions. This study established strong evidence that peat microbes and water table fluctuation can be an excellent tool for hydrocarbon removal and its control in peatlands.
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Agua Subterránea , Microbiota , Contaminación Ambiental , Humanos , Hidrocarburos , SueloRESUMEN
An amendment to this paper has been published and can be accessed via the original article.