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OBJECTIVE: The microbiome contributes to the pathogenesis of inflammatory bowel disease (IBD) but the relative contribution of different lifestyle and environmental factors to the compositional variability of the gut microbiota is unclear. DESIGN: Here, we rank the size effect of disease activity, medications, diet and geographic location of the faecal microbiota composition (16S rRNA gene sequencing) in patients with Crohn's disease (CD; n=303), ulcerative colitis (UC; n = 228) and controls (n=161), followed longitudinally (at three time points with 16 weeks intervals). RESULTS: Reduced microbiota diversity but increased variability was confirmed in CD and UC compared with controls. Significant compositional differences between diseases, particularly CD, and controls were evident. Longitudinal analyses revealed reduced temporal microbiota stability in IBD, particularly in patients with changes in disease activity. Machine learning separated disease from controls, and active from inactive disease, when consecutive time points were modelled. Geographic location accounted for most of the microbiota variance, second to the presence or absence of CD, followed by history of surgical resection, alcohol consumption and UC diagnosis, medications and diet with most (90.3%) of the compositional variance stochastic or unexplained. CONCLUSION: The popular concept of precision medicine and rational design of any therapeutic manipulation of the microbiota will have to contend not only with the heterogeneity of the host response, but also with widely differing lifestyles and with much variance still unaccounted for.
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Microbioma Gastrointestinal , Enfermedades Inflamatorias del Intestino/microbiología , Estilo de Vida , Canadá , Dieta , Femenino , Geografía , Humanos , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Irlanda , Estudios Longitudinales , Aprendizaje Automático , Masculino , Persona de Mediana Edad , Encuestas y CuestionariosRESUMEN
Improvements in bacterial culturing and DNA sequencing techniques have revealed a diverse, and hitherto unknown, urinary tract microbiome (urobiome). The potential role of this microbial community in contributing to health and disease, particularly in the context of urinary tract infections (UTIs) is of significant clinical importance. However, while several studies have confirmed the existence of a core urobiome, the role of its constituent microbes is not yet fully understood, particularly in the context of health and disease. Herein, we review the current state of the art, concluding that the urobiome represents an important component of the body's innate immune defences, and a potentially rich resource for the development of alternative treatment and control strategies for UTIs.
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Infecciones por Escherichia coli , Infecciones Urinarias , Sistema Urinario , Escherichia coli Uropatógena , Humanos , Infecciones Urinarias/tratamiento farmacológicoRESUMEN
Cryptosporidiosis remains the leading protozoan induced cause of diarrhoea-associated mortality worldwide. Cryptosporidium hominis, the anthroponotically transmitted species within the Cryptosporidium genus, contributes significantly to the global burden of infection, accounting for the majority of clinical cases in many countries. This study applied high resolution melting analysis, a post-real-time PCR application, to the differentiation of six globally prevalent C. hominisgp60-subtypes. This novel method targeted three microsatellite, tandem repeat containing genetic markers, gp60, the genetic marker upon which current Cryptosporidium subtype nomenclature is based, MSB, and MSE, by which to differentiate between C. hominis isolates. This multi-locus approach successfully differentiated between all six C. hominisgp60-subtypes studied, some of which, such as IbA10G2, are known to exhibit global ubiquity. Thus, this method has the potential to be universally employed as a sensitive, cost effective and highly reproducible means to rapidly differentiate between C. hominisgp60-subtypes. Such a method would be of particular utility in epidemiological studies and outbreak scenarios, providing cost effective, clinically accessible alternative to DNA sequencing. The success of this preliminary study also supports further analysis of an expanded C. hominisgp60-subtype range and the potential expansion of the multi-locus panel in order to improve the discriminatory power of this approach.
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Cryptosporidium/genética , Parasitología/métodos , Criptosporidiosis/parasitología , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , ADN Protozoario/genética , Heces/parasitología , Marcadores Genéticos , Genotipo , Humanos , Tipificación de Secuencias Multilocus , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
The objective of this study was to determine whether response to selection for carcass weight (CW), conformation (CC) and fat (CF), and the association between heterosis and carcass performance varied by herd environment in cattle. Following edits, carcass information was available for 4,616,761 cattle, of which the majority were some crossbred combination of the following breeds: Angus, Aubrac, Belgian Blue, Blonde d'Aquitaine, Charolais, Hereford, Holstein-Friesian, Jersey, Limousin, Saler, Shorthorn and Simmental. Herd environment was defined separately for each carcass trait using herd solutions outputted from carcass trait genetic evaluations. A total of 3,859 herds were stratified, for each trait, into one of five strata based on their corresponding percentile herd solution rank, with the response to selection and the effect of heterosis then estimated within each stratum. The response in CW and CC from selection on the respective estimated breeding values (EBV) increased between the lowest (0.71 kg and 0.89 CC score increase per unit increase in the respective EBV) and highest (0.99 kg and 1.25 CC score increase per unit increase in the respective EBV) corresponding herd stratum. The response in CF from selection on CF EBV, however, reduced between the lowest and highest CF herd stratum (respective increases of 0.93 and 0.83 CF scores per unit increase in CF EBV). In addition, the effect of a unit increase in heterosis coefficient on CW, CC and CF also varied by herd stratum. Furthermore, results (i.e. the area under relative operating characteristic curves) from the present study demonstrated that the response to selection and heterosis effects estimated for the different herd stratum can be used, along with EBVs and the herd solutions themselves, to improve the accuracy of phenotypic predictions. Results from the present study could help producers to make more informed breeding decisions that are bespoke to their herd.
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Vigor Híbrido , Animales , Bovinos/genética , FenotipoRESUMEN
Reported incidence rates of cryptosporidiosis in Ireland are consistently among the highest in Europe. Despite the national prevalence of this enteric parasite and the compulsory nature of incidence surveillance and reporting, in-depth analyses seeking to genotype clinical isolates of Cryptosporidium on an intra-species level are rarely undertaken in Ireland. This molecular epidemiology study of 163 clinical Cryptosporidium isolates was conducted in Southern Ireland, from 2015 to 2018, in order to ascertain population subtype heterogeneity. Analysis was conducted via real-time PCR amplification and gp60 gene sequencing, which successfully determined the subtype designation of 149 of the 163 (91.4%) tested isolates. Overall, 12 C. parvum and five C. hominis subtypes were identified, with the incidence of the regionally predominant C. parvum species found to primarily occur during springtime months, while C. hominis incidence was largely confined to late summer and autumnal months. Additionally, one C. parvum and four C. hominis subtypes were newly reported by this study, having not been previously identified in clinical or livestock infection in Ireland. Overall, these data give insight into the diversification of the Cryptosporidium population and emergent subtypes, while also allowing comparisons to be made with clinical epidemiological profiles reported previously in Ireland and elsewhere.
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Criptosporidiosis/parasitología , Cryptosporidium/clasificación , Zoonosis/parasitología , Animales , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Cryptosporidium parvum/clasificación , Cryptosporidium parvum/genética , Cryptosporidium parvum/aislamiento & purificación , ADN Protozoario/aislamiento & purificación , Heces/parasitología , Gastroenteritis/parasitología , Glicoproteínas/química , Glicoproteínas/genética , Humanos , Incidencia , Irlanda/epidemiología , Estudios Longitudinales , Prevalencia , Proteínas Protozoarias/química , Proteínas Protozoarias/genética , ARN Ribosómico 18S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios Retrospectivos , Estaciones del Año , Alineación de SecuenciaRESUMEN
Objectives: This study considers susceptibility test results obtained over a 6 month period for Enterobacteriaceae that caused urinary tract infections (UTIs) in the Cork region of Ireland and uses these results to examine the suitability of Irish empirical treatment guidelines. Patients and methods: UTI-causing Enterobacteriaceae isolates were analysed using EUCAST guidelines to determine resistance to a set of commonly prescribed antimicrobial agents, i.e. ampicillin, amoxicillin/clavulanate, cefalexin, ciprofloxacin, nitrofurantoin and trimethoprim. Patients were categorized by age and patient type, based on origin (hospital inpatients, patients in long-term care facilities and all other non-hospitalized patients). In total, 8999 test results were analysed using the IBM Cognos Analytics Series 7 interrogation tool and Microsoft Office Excel. Results: A variety of resistance patterns were observed. Only one antimicrobial agent, nitrofurantoin, demonstrated a resistance rate of less than 20% for all patient categories considered. Conclusions: Previous studies determined that a resistance rate of >20% renders an antimicrobial agent unsuitable for use as an empirical treatment option. This study demonstrated that this resistance rate is exceeded in many cases, potentially rendering some antimicrobial agents unsuitable for use as empirical treatment. We suggest that the focus on susceptibility when producing surveillance data to create empirical treatment guidelines may inadvertently camouflage resistance rates. The findings of this study highlight the need for laboratory-guided treatment of UTIs and ideally a pre-emptive sample should be obtained for laboratory investigation prior to commencement of antimicrobial therapy.
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Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana , Infecciones por Enterobacteriaceae/tratamiento farmacológico , Enterobacteriaceae/efectos de los fármacos , Guías de Práctica Clínica como Asunto , Infecciones Urinarias/tratamiento farmacológico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Enterobacteriaceae/aislamiento & purificación , Infecciones por Enterobacteriaceae/microbiología , Femenino , Humanos , Lactante , Recién Nacido , Irlanda , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , Infecciones Urinarias/microbiología , Adulto JovenRESUMEN
Synthetic biology, the design and synthesis of synthetic biological systems from DNA to whole cells, has provided us with the ultimate tools for space exploration and colonisation. Herein, we explore some of the most significant advances and future prospects in the field of synthetic biology, in the context of astrobiology and terraforming.
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Exobiología/tendencias , Biología Sintética/tendencias , ADN/síntesis química , ADN/genética , Exobiología/métodos , Vuelo Espacial , Biología Sintética/métodosRESUMEN
BACKGROUND: The field of diagnostics continues to advance rapidly with a variety of novel approaches, mainly dependent upon high technology platforms. Nonetheless much diagnosis, particularly in developing countries, still relies upon traditional methods such as microscopy. Biological material, particularly nucleic acids, on archived glass slides is a potential source of useful information both for diagnostic and epidemiological purposes. There are significant challenges faced when examining archived samples in order that an adequate amount of amplifiable DNA can be obtained. Herein, we describe a model system to detect low numbers of bacterial cells isolated from glass slides using (laser capture microscopy) LCM coupled with PCR amplification of a suitable target. RESULTS: Mycobacterium smegmatis was used as a model organism to provide a proof of principle for a method to recover bacteria from a stained sample on a glass slide using a laser capture system. Ziehl-Neelsen (ZN) stained cells were excised and catapulted into tubes. Recovered cells were subjected to DNA extraction and pre-amplified with multiple displacement amplification (MDA). This system allowed a minimum of 30 catapulted cells to be detected following a nested real-time PCR assay, using rpoB specific primers. The combination of MDA and nested real-time PCR resulted in a 30-fold increase in sensitivity for the detection of low numbers of cells isolated using LCM. CONCLUSIONS: This study highlights the potential of LCM coupled with MDA as a tool to improve the recovery of amplifiable nucleic acids from archived glass slides. The inclusion of the MDA step was essential to enable downstream amplification. This platform should be broadly applicable to a variety of diagnostic applications and we have used it as a proof of principle with a Mycobacterium sp. model system.
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ADN Bacteriano/aislamiento & purificación , Microscopía Confocal/métodos , Mycobacterium smegmatis/aislamiento & purificación , ADN Bacteriano/genética , Vidrio/análisis , Humanos , Infecciones por Mycobacterium no Tuberculosas/diagnóstico , Infecciones por Mycobacterium no Tuberculosas/microbiología , Mycobacterium smegmatis/clasificación , Mycobacterium smegmatis/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y Especificidad , Coloración y Etiquetado/instrumentaciónRESUMEN
Nutritionally, the first 1,000 days of an infant's life - from conception to two years - has been identified as a highly influential period, during which lasting health can be achieved. Significant evidence links patterns of infant feeding to both short and long-term health outcomes, many of which can be prevented through nutritional modifications. Recommended globally, breastfeeding is recognised as the gold standard of infant nutrition; providing key nutrients to achieve optimal health, growth and development, and conferring immunologic protective effects against disease. Nevertheless, infant formulas are often the sole source of nutrition for many infants during the first stage of life. Producers of infant formula strive to supply high quality, healthy, safe alternatives to breast milk with a comparable balance of nutrients to human milk imitating its composition and functional performance measures. The concept of 'nutritional programming', and the theory that exposure to specific conditions, can predispose an individual's health status in later life has become an accepted dictum, and has sparked important nutritional research prospects. This review explores the impact of early life nutrition, specifically, how different feeding methods affect health outcomes.
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Fenómenos Fisiológicos Nutricionales del Lactante , Alimentación con Biberón/efectos adversos , Alimentación con Biberón/métodos , Lactancia Materna/métodos , Humanos , Lactante , Fórmulas Infantiles/química , Fórmulas Infantiles/normas , Recién Nacido , Valor NutritivoRESUMEN
Applying 21st century technology to the design and development of a hypothetical extra-terrestrial colonisation programme, we reimagine 'directed panspermia' from the perspective of Crick and Orgel's 'technological society', 44 years after the publication of their original landmark paper.
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The recent paradigm shift in infectious disease diagnosis from culture-based to molecular-based approaches is exemplified in the findings of a national study assessing the detection of verotoxigenic Escherichia coli infections in Ireland. The methodologic changes have been accompanied by a dramatic increase in detections of non-O157 verotoxigenic E. coli serotypes.
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Infecciones por Escherichia coli/diagnóstico , Infecciones por Escherichia coli/microbiología , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Infecciones por Escherichia coli/epidemiología , Humanos , Irlanda/epidemiología , Tipificación Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa , Serogrupo , Serotipificación , Toxina Shiga/genéticaRESUMEN
Existing at the interface of science and engineering, synthetic biology represents a new and emerging field of mainstream biology. However, there also exists a counterculture of Do-It-Yourself biologists, citizen scientists, who have made significant inroads, particularly in the design and development of new tools and techniques. Herein, I review the development and convergence of synthetic biology's mainstream and countercultures.
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Investigación Biomédica , Biotecnología , Biología Sintética , Contención de Riesgos Biológicos , HumanosRESUMEN
The D-It-Yourself Biology (DIYBio) movement has gained significant traction in recent years; morphing from counter culture to an essential component of biology's future development.
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Weissella is a genus of lactic acid bacteria (LAB) consisting of species formerly included in the Leuconostoc paramesenteroides group. Similar to other LAB, they are commonly found in fermented foods but have also been isolated from environmental and human samples. Currently there are 20 recognized species. Herein, three Weissella cibaria genomes were sequenced using Illumia Mi-Seq and Roche 454 technologies. Annotation was performed using the Prokka and JGI IMG pipelines. A thorough analysis of the genomics of the W. cibaria strains was performed, in addition to brief comparative analyses of the genus Weissella as a whole. Genomic sequence data from the newly sequenced W. cibaria strains and data available in GenBank for other Weissella strains was used (nâ=â10; four Weissella cibaria, one Weissella ceti, one Weissella confusa, one Weissella halotolerans, two Weissella koreensis and one Weissella paramesenteroides). The genomes had sizes varying from 1.3 to 2.4 Mb. DNA G+C contents ranged from 35 to 45 mol%. The core- and pan-proteome at genus and species levels were determined. The genus pan-proteome was found to comprise 4712 proteins. Analysis of the four W. cibaria genomes indicated that the core-proteome, consisting of 729 proteins, constitutes 69â% of the species pan-proteome. This large core-set may explain the divergent niches in which this species has been found. In W. cibaria, in addition to a number of phosphotransferase systems conferring the ability to assimilate plant-associated polysaccharides, an extensive proteolytic system was identified.
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Genómica , Carácter Cuantitativo Heredable , Weissella/genética , Weissella/metabolismo , Adaptación Biológica , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Bacteriófagos , Metabolismo Energético , Genómica/métodos , Secuencias Repetitivas Esparcidas , Datos de Secuencia Molecular , Consumo de Oxígeno , Filogenia , Proteoma , Proteómica , ARN Ribosómico 16S/genética , Alineación de Secuencia , Weissella/virologíaRESUMEN
The recent exponential growth of genomic databases has resulted in the common task of sequence alignment becoming one of the major bottlenecks in the field of computational biology. It is typical for these large datasets and complex computations to require cost prohibitive High Performance Computing (HPC) to function. As such, parallelised solutions have been proposed but many exhibit scalability limitations and are incapable of effectively processing "Big Data" - the name attributed to datasets that are extremely large, complex and require rapid processing. The Hadoop framework, comprised of distributed storage and a parallelised programming framework known as MapReduce, is specifically designed to work with such datasets but it is not trivial to efficiently redesign and implement bioinformatics algorithms according to this paradigm. The parallelisation strategy of "divide and conquer" for alignment algorithms can be applied to both data sets and input query sequences. However, scalability is still an issue due to memory constraints or large databases, with very large database segmentation leading to additional performance decline. Herein, we present Hadoop Blast (HBlast), a parallelised BLAST algorithm that proposes a flexible method to partition both databases and input query sequences using "virtual partitioning". HBlast presents improved scalability over existing solutions and well balanced computational work load while keeping database segmentation and recompilation to a minimum. Enhanced BLAST search performance on cheap memory constrained hardware has significant implications for in field clinical diagnostic testing; enabling faster and more accurate identification of pathogenic DNA in human blood or tissue samples.
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Bases de Datos Genéticas , Genómica/métodos , Alineación de Secuencia/métodos , Programas Informáticos , Algoritmos , Metodologías Computacionales , Bases de Datos Factuales , Técnicas Genéticas , Humanos , InternetRESUMEN
An investigation of the prevalence of Campylobacter ureolyticus in a variety of animals led to the identification of the strain CIT 045(T), in the faeces of captive lion-tailed macaques (Macaca silenus). Originally, believed to be Campylobacter ureolyticus based on the colony morphology and positive urease test, analysis of 16S rRNA and hsp60 gene sequences of this isolate revealed that the strain differs significantly from other species of the genus Campylobacter described to date. Species-specific primers for 16S rRNA and hsp60 genes were designed and used to identify two additional strains isolated from faeces samples from other macaques. Nucleotide sequence analysis of the 16S rRNA and hsp60 genes revealed ≤95% and ≤82 â% sequence similarity to recognized species of the genus Campylobacter respectively. All three isolates formed a distinct group within the genus Campylobacter based on their 16S rRNA and hsp60 sequences and matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) profiles. The unique species status was further supported by phenotypic characteristics of the isolates. All isolates were found to be oxidase-, catalase- and urease-positive, they grew well at 37 °C and 42 °C and produced H2S on TSI (triple-sugar iron) and SIM (sulfide indole motility) media. The name Campylobacter corcagiensis sp. nov. is proposed for this novel species, with the strain CIT 045(T) as the type strain CIT 045(T) (â=âLMG 27932(T), CCUG 64942(T)).
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Campylobacter/clasificación , Heces/microbiología , Macaca/microbiología , Filogenia , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Campylobacter/genética , Campylobacter/aislamiento & purificación , ADN Bacteriano/genética , Genes Bacterianos , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
During the early stages of the COVID-19 pandemic, we called for mandatory public masking to 'flatten the curve'. We helped formulate a national standard (SWiFT 19) for barrier facemasks, and, using a novel laser-based approach, we determined that mask efficacy is dependent on both fabric and fit; with both variables being inversely related. Herein, we take a retrospective view of the role of masks during the pandemic and surmise that, on the balance of evidence to date, masks were effective at stemming the spread of SARS-CoV-2 and may well be an effective early control strategy for potential future respiratory pandemics.
Face coverings, which cover the nose and mouth, are a means of preventing infections that travel in the air. These include viruses such as SARS-CoV-2, which causes COVID-19. Face coverings, or masks, played a key role during the COVID-19 pandemic by reducing person-to-person spread of the virus. The key features of a mask that make it effective are the material from which it is made and how closely the mask fits the face. A loosely fitting mask, for example, will lead to gaps around the nose and cheeks through which droplets can escape. A better fitting mask will have less leakage. Masks made from light single-layer material is less able to prevent droplet penetration than thicker, multi-layered fabric. Properly fashioned and fitted face masks are an effective means of slowing the spread of infections that travel in the air.
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COVID-19 , Máscaras , SARS-CoV-2 , COVID-19/prevención & control , COVID-19/epidemiología , COVID-19/transmisión , Humanos , Pandemias/prevención & control , Estudios RetrospectivosRESUMEN
Neuronal development is a highly regulated mechanism that is central to organismal function in animals. In humans, disruptions to this process can lead to a range of neurodevelopmental phenotypes, including Schizophrenia (SCZ). SCZ has a significant genetic component, whereby an individual with an SCZ affected family member is eight times more likely to develop the disease than someone with no family history of SCZ. By examining a combination of genomic, transcriptomic and epigenomic datasets, large-scale 'omics' studies aim to delineate the relationship between genetic variation and abnormal cellular activity in the SCZ brain. Herein, we provide a brief overview of some of the key omics methods currently being used in SCZ research, including RNA-seq, the assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) and high-throughput chromosome conformation capture (3C) approaches (e.g., Hi-C), as well as single-cell/nuclei iterations of these methods. We also discuss how these techniques are being employed to further our understanding of the genetic basis of SCZ, and to identify associated molecular pathways, biomarkers, and candidate drug targets.
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Esquizofrenia , Animales , Humanos , Esquizofrenia/genética , Cromatina/genética , Cromatina/metabolismo , Secuenciación de Inmunoprecipitación de Cromatina , Encéfalo/metabolismo , Secuenciación de Nucleótidos de Alto RendimientoRESUMEN
Molecular diagnostics has the potential to revolutionise the field of clinical microbiology. Microbial identification and nomenclature have, for too long, been restricted to phenotypic characterisation. However, this species-level view fails to wholly account for genetic heterogeneity, a result of lateral gene transfer, mediated primarily by mobile genetic elements. This genetic promiscuity has helped to drive virulence development, stress adaptation, and antimicrobial resistance in several important bacterial pathogens, complicating their detection and frustrating our ability to control them. We argue that, as clinical microbiologists at the front line, we must embrace the molecular technologies that allow us to focus specifically on the genetic elements that cause disease rather than the bacterial species that express them. This review focuses on the evolution of microbial taxonomy since the introduction of molecular sequencing, the role of mobile genetic elements in antimicrobial resistance, the current and emerging assays in clinical laboratories, and the comparison of phenotypic versus genotypic analyses. In essence, it is time now to refocus from species to genes as part of a new diagnostic paradigm.
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Background: The role of the urobiome in health and disease remains an understudied area compared to the rest of the human microbiome. Enhanced culturing techniques and next-generation sequencing technologies have identified the urobiome as an untapped source of potentially novel antimicrobials. The aim of this study was to screen the urobiome for genes encoding bacteriocin production. Methods: The genomes of 181 bacterial urobiome isolates were screened in silico for the presence of bacteriocin gene clusters using the bacteriocin mining tool BAGEL4 and secondary metabolite screening tool antiSMASH7. Results: From these isolates, an initial 263 areas of interest were identified, manually annotated, and evaluated for potential bacteriocin gene clusters. This resulted in 32 isolates containing 80 potential bacteriocin gene clusters, of which 72% were identified as class II, 13.75% as class III, 8.75% as class I, and 5% as unclassified bacteriocins. Conclusion: Overall, 53 novel variants were discovered, including nisin, gassericin, ubericin, and colicins.