Comparative genomics of Campylobacter concisus:Analysis of clinical strains reveals genome diversity and pathogenic potential

We thank members of the GI Research Team for discussions and advice. The authors thank Brennan Martin and the Centre for Genome Enabled Biology and Medicine for Illumina sequencing and useful discussions. This work was supported by a Fulbright Scholarship to G.L.H., an NHS Grampian Endowment grant fund to I.M. and G.L.H., a CSO clinical academic fellowship to R.H. (CAF/08/01). R.H. is supported by an NHS Research Scotland Career Researcher Fellowship. This work was generously supported by the Catherine McEwan Foundation. Sequence deposition The C. concisus raw sequencing reads and genome assemblies are freely available from the EMBL-EBI ENA under the study Accession PRJEB22351.Peer reviewedPublisher PD

Novel Campylobacter concisus lipooligosaccharide is a determinant of inflammatory potential and virulence

This work was supported in part by Department of Veterans Affairs Merit Review award BX000727 (to G.A.J.). The authors also acknowledge National Institutes of Health National Center for Research Resources Shared Instrumentation Grant S10 RR029446 (to H. E. Witkowska) for acquisition of the Synapt G2 high definition mass spectrometer, which is located at the University of California, San Francisco Sandler-Moore Mass Spectrometry Core Facility and supported by the Sandler Family Foundation, the Gordon and Betty Moore Foundation, National Institutes of Health/National Cancer Institute Cancer Center Support Grant P30 CA082103, and the Canary Foundation. G.A.J. is a recipient of the Senior Research Career Scientist award from the Department of Veterans Affairs. R.H. is funded by a Career Researcher Fellowship from NHS Research Scotland. The BISCUIT study was funded by a Clinical Academic Training Fellowship from the Chief Scientist Office (CAF/08/01). This is paper number 116 from the Center for Immunochemistry. The contents of this article do not represent the views of the Department of Veterans Affairs or the United States Government. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. K.B. acknowledges funding from the Child Health Research Charitable Incorporated Organisation and the Bogue Fellowship for travel. The authors declare that they have no conflicts of interest with the contents of this article.Peer reviewe

Type II and type IV toxin–antitoxin systems show different evolutionary patterns in the global Klebsiella pneumoniae population

The Klebsiella pneumoniae species complex includes important opportunistic pathogens which have become public health priorities linked to major hospital outbreaks and the recent emergence of multidrug-resistant hypervirulent strains. Bacterial virulence and the spread of multidrug resistance have previously been linked to toxin-antitoxin (TA) systems. TA systems encode a toxin that disrupts essential cellular processes, and a cognate antitoxin which counteracts this activity. Whilst associated with the maintenance of plasmids, they also act in bacterial immunity and antibiotic tolerance. However, the evolutionary dynamics and distribution of TA systems in clinical pathogens are not well understood. Here we present a comprehensive survey and description of the diversity of TA systems in 259 clinically relevant genomes of K. pneumoniae. We show that TA systems are highly prevalent with a median of 20 loci per strain. Importantly, these toxins differ substantially in their distribution patterns and in their range of cognate antitoxins. Classification along these properties suggests different roles of TA systems and highlights the association and co-evolution of toxins and antitoxins

Molecular epidemiology and antimicrobial resistance of outbreaks of Klebsiella pneumoniae clinical mastitis in Chinese dairy farms

Klebsiella pneumoniae is an opportunistic pathogen that causes serious infections in humans and animals. However, the availability of epidemiological information on clinical mastitis due to K. pneumoniae is limited. To acquire new information regarding K. pneumoniae mastitis, data were mined about K. pneumoniae strains on dairy cattle farms (farms A to H) in 7 Chinese provinces in 2021. Hypermucoviscous strains of K. pneumoniae were obtained by the string test. MICs of antimicrobial agents were determined via the broth microdilution method. Ten antimicrobial resistance genes and virulence genes were identified by PCR. The prevalence of K. pneumoniae was 35.91% (65/181), and 100% of the bacteria were sensitive to enrofloxacin. Nine antimicrobial resistance genes and virulence genes were identified and compared among farms. The hypermucoviscous phenotype was present in 94.44% of isolates from farm B, which may be a function of the rmpA virulence gene. Based on these data, the multidrug-resistant strains SD-14 and HB-21 were chosen and sequenced. Genotypes were assayed for K. pneumoniae isolates from different countries and different hosts using multilocus sequence typing (MLST). Ninety-four sequence types (STs) were found, and 6 STs present a risk for spreading in specific regions. Interestingly, ST43 was observed in bovine isolates for the first time. Our study partially reveals the current distribution characteristics of bovine K. pneumoniae in China and may provide a theoretical basis for the prevention and treatment of bovine K. pneumoniae mastitis

Comparative analysis of amplicon and metagenomic sequencing methods reveals key features in the evolution of animal metaorganisms

Background The interplay between hosts and their associated microbiome is now recognized as a fundamental basis of the ecology, evolution, and development of both players. These interdependencies inspired a new view of multicellular organisms as “metaorganisms.” The goal of the Collaborative Research Center “Origin and Function of Metaorganisms” is to understand why and how microbial communities form long-term associations with hosts from diverse taxonomic groups, ranging from sponges to humans in addition to plants. Methods In order to optimize the choice of analysis procedures, which may differ according to the host organism and question at hand, we systematically compared the two main technical approaches for profiling microbial communities, 16S rRNA gene amplicon and metagenomic shotgun sequencing across our panel of ten host taxa. This includes two commonly used 16S rRNA gene regions and two amplification procedures, thus totaling five different microbial profiles per host sample. Conclusion While 16S rRNA gene-based analyses are subject to much skepticism, we demonstrate that many aspects of bacterial community characterization are consistent across methods. The resulting insight facilitates the selection of appropriate methods across a wide range of host taxa. Overall, we recommend single- over multi-step amplification procedures, and although exceptions and trade-offs exist, the V3 V4 over the V1 V2 region of the 16S rRNA gene. Finally, by contrasting taxonomic and functional profiles and performing phylogenetic analysis, we provide important and novel insight into broad evolutionary patterns among metaorganisms, whereby the transition of animals from an aquatic to a terrestrial habitat marks a major event in the evolution of host-associated microbial composition

Desarrollo y despliegue de un workflow para el análisis genómico de Campylobacter Jejuni

Trabajo de fin de máster en Bioinformática y Biología ComputacionalEl auge de las nuevas técnicas de secuenciación masiva está generando una necesidad de análisis de gran cantidad de datos que provoca el acercamiento a la bioinformática de nuevos grupos de investigación antes ajenos a ella. Grupos que solían encargar los análisis de sus datos a otros grupos de investigación están comenzando a desarrollar sus propios workflows para tratar los datos personalmente. En este contexto, el grupo de investigación Tecnofood de la Universidad de Burgos, con una larga trayectoria en investigación sobre las fuentes de contaminación en la cadena alimentaria, ha decidido desarrollar un workflow destinado al análisis genómico de la bacteria Campylobacter jejuni. Este trabajo estudia su proceso de creación, orientado a establecer una primera herramienta accesible al grupo con la que tendrán la posibilidad de realizar análisis, tanto de las cepas de esta bacteria, como de otras similares en un futuro. La interfaz gráfica de la infraestructura utilizada para su soporte, Galaxy, hace posible que el workflow pueda sufrir modificaciones de manera sencilla en un futuro. Esto permitirá dar lugar a variaciones del workflow sin necesidad de un nuevo desarrollo poco accesible a usuarios ajenos a la programación. El sistema se ha desplegado sobre un contenedor Docker con la intención de ahorrar trabajo al usuario evitando la instalación y configuración de un entorno apropiado. El workflow consta de 6 utilidades principales que resuelven los aspectos de filtrado de calidad (Trimmomatic y Prinseq), ensamblado (SPAdes), etiquetado (Prokka), análisis de resistencia a antibióticos (ABRicate) y análisis pangenómico (Roary). Finalmente, el funcionamiento de este workflow se ha puesto a prueba utilizando cuarenta y seis cepas de Campylobacter jejuni, secuenciadas con un equipo Illumina, procedentes de un matadero del norte de España

Caracterización fisiológica y genómica de dos cepas nativas del género Shewanella con potencial biodegradador de colorantes azoicos

La industria textil libera al ambiente grandes cantidades de aguas residuales que, entre otros varios contaminantes, contienen colorantes azo en su composición, los cuales son compuestos tóxicos, carcinogénicos y mutagénicos que afectan negativamente la vida acuática y la calidad del agua. El presente estudio tuvo por objetivo caracterizar a nivel fisiológico y genómico cepas nativas con potencial de degradación de colorantes azo obtenidas de un efluente de industria textil de Lima, Perú. Se aplicó el método de la microplaca a doce aislados bacterianos para su selección en base a su respuesta positiva a la decoloración de tres colorantes azo: Azul directo 71, Rojo remazol RGB y Amarillo Oro Remazol RGB. La actividad decolorante de las cepas seleccionadas fue evaluada mediante espectrofotometría UV-Visible en medio ZZ con el colorante respectivo a 100 ppm y en condiciones de microaerofilia durante 24 horas. El secuenciamiento de los genomas se realizó utilizando la tecnología HiSeq 2500 de Illumina, el ensamblaje de novo con el software SPAdes, la extensión de los contigs y reparación de gaps a través de los programas ABACAS e IMAGE y la anotación con Prokka. Las dos cepas con mejor eficiencia de degradación correspondieron a Shewanella sp. LC-2 y Shewanella sp. LC-6, ambas identificadas mediante análisis filogenómico resultando muy cercanas al grupo filogenético de Shewanella sp. FDAARGOS_354. Las cepas mostraron una actividad decolorante frente a Azul Directo 71, Rojo Remazol y Amarillo Remazol con porcentajes de 94.42, 94.79, 91.67 para Shewanella sp. LC-2 y 94.37, 94.92, 83.24 para Shewanella sp. LC-6 a las 24 h. Asimismo, los genomas de Shewanella sp. LC-2 y LC-6 revelaron la presencia de genes que codifican azorreductasas dependientes de NADH, peroxidasas decoloradoras de tinte (DyPs), genes implicados en la desaminación, asimilación de sulfatos, reducción de nitratos y metales pesados, como también en la degradación de benzoatos, catecol y gentisato. Se concluye que las cepas Shewanella LC-2 y LC-6 tienen una actividad metabólica eficaz para la decoloración de Azul directo 71, Rojo remazol RGB y Amarillo Oro Remazol RGB, presentan versatilidad catabólica y son de potencial aplicación en la biorremediación de aguas residuales textiles.Universidad Nacional Mayor de San Marcos (Lima). Vicerrectorado de Investigación y PosgradoPerú. Ministerio de la Producción. Programa Nacional de Innovación para la Competitividad y Productividad (Innóvate Perú). Fondo para la Innovación, la Ciencia y la Tecnología (FINCyT)Tesi

Pangenome insights into the diversification and disease specificity of worldwide Xanthomonas outbreaks

The bacterial genus Xanthomonas is responsible for disease outbreaks in several hundred plant species, many of them economically important crops. In the era of next-generation sequencing, thousands of strains from this genus have now been sequenced as part of isolated studies that focus on outbreak characterization, host range, diversity, and virulence factor identification. However, these data have not been synthesized and we lack a comprehensive phylogeny for the genus, with some species designations in public databases still relying on phenotypic similarities and representative sequence typing. The extent of genetic cohesiveness among Xanthomonas strains, the distribution of virulence factors across strains, and the impact of evolutionary history on host range across the genus are also poorly understood. In this study, we present a pangenome analysis of 1,910 diverse Xanthomonas genomes, highlighting their evolutionary relationships, the distribution of virulence-associated genes across strains, and rates of horizontal gene transfer. We find a number of broadly conserved classes of virulence factors and considerable diversity in the Type 3 Secretion Systems (T3SSs) and Type 3 Secreted Effector (T3SE) repertoires of different Xanthomonas species. We also use these data to re-assign incorrectly classified strains to phylogenetically informed species designations and find evidence of both monophyletic host specificity and convergent evolution of phylogenetically distant strains to the same host. Finally, we explore the role of recombination in maintaining genetic cohesion within the Xanthomonas genus as a result of both ancestral and recent recombination events. Understanding the evolutionary history of Xanthomonas species and the relationship of key virulence factors with host-specificity provides valuable insight into the mechanisms through which Xanthomonas species shift between hosts and will enable us to develop more robust resistance strategies against these highly virulent pathogens