Stephen Mark Edward Fordham, Magdalena Barrow, Anna Mantzouratou, Elizabeth Sheridan
{"title":"Genomic analyses of an Escherichia coli and Klebsiella pneumoniae urinary tract co-infection using long-read nanopore sequencing","authors":"Stephen Mark Edward Fordham, Magdalena Barrow, Anna Mantzouratou, Elizabeth Sheridan","doi":"10.1002/mbo3.1396","DOIUrl":"https://doi.org/10.1002/mbo3.1396","url":null,"abstract":"<p><i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> isolates presenting with the same antimicrobial susceptibility profile were recovered from the same catheter sample of urine (CSU). Both strains were recovered from a patient with a long-standing indwelling urinary catheter. Each isolate had its DNA extracted following culture. Nanopore long-read sequencing was used to build the plasmids and chromosomes from each strain to closure to discern the potential horizontal propagation of resistance-encoding plasmids and the relationship between resistance genes and insertion sequences. Plasmids derived from resistance strains in the urinary microbiota remain poorly characterized. The same 11 antimicrobial resistance (AMR) genes were found in plasmids from each strain. The 185,239-bp FIB(K) pKBM1, from the <i>K. pneumoniae</i> strain, additionally encoded the five AMR genes: <i>sul2, strA, strB, bla</i><sub>TEM-1B</sub>, and <i>bla</i><sub>CTX-M-15</sub>. A multimeric array of AMR genes and IS<i>26</i> insertion sequences were found in the plasmids from both isolates. Both plasmids from each isolate were similar. Horizontal transfer of plasmids, followed by subsequent plasmid rearrangement, is likely to have occurred during infection. Furthermore, the resistance region in the plasmids shared similarity against the internationally prevalent plasmid, pKPN3-307_typeA, commonly identified in <i>K. pneumoniae</i> ST307. Biofilm formation in catheterized patients may allow close cell contact between strains. Horizontal propagation of resistance genes may occur, leading to polymicrobial infections.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1396","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139488617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Laura Marquardt, Federica Andreoni, Mathilde Boumasmoud, Tiziano A. Schweizer, Dorothea M. Heuberger, Elena Parietti, Sanne Hertegonne, Jana Epprecht, Dario Mattle, Anna K. Raez, Ewerton Marques-Maggio, Reto A. Schuepbach, Barbara Hasse, Srikanth Mairpady-Shambat, Silvio D. Brugger, Annelies S. Zinkernagel
{"title":"Group A Streptococcus strains causing meningitis without distinct invasive phenotype","authors":"Laura Marquardt, Federica Andreoni, Mathilde Boumasmoud, Tiziano A. Schweizer, Dorothea M. Heuberger, Elena Parietti, Sanne Hertegonne, Jana Epprecht, Dario Mattle, Anna K. Raez, Ewerton Marques-Maggio, Reto A. Schuepbach, Barbara Hasse, Srikanth Mairpady-Shambat, Silvio D. Brugger, Annelies S. Zinkernagel","doi":"10.1002/mbo3.1394","DOIUrl":"10.1002/mbo3.1394","url":null,"abstract":"<p>Group A streptococcal (GAS; aka <i>Streptococcus pyogenes</i>) meningitis is a fulminant disease associated with high morbidity and mortality. To elucidate the mechanisms underlying the invasiveness of GAS in meningitis, we compared GAS isolates derived from five cases of meningitis to otitis and colonizing isolates. We did not observe differences in adherence to and invasion of human brain microvascular endothelial cells, virulence factors activity, or barrier disruption. Whole genome sequencing did not reveal particular invasiveness traits. Most patients previously suffered from otitis media suggesting that meningitis likely resulted from a continuous spread of the infection rather than being attributable to changes in the pathogen's virulence.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1394","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139094480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lesley Sitter, Marion Schoof, Travis R. Glare, Murray P. Cox, Peter C. Fineran, Paul P. Gardner, Mark R. H. Hurst
{"title":"Serratia-based toxin cluster elements are associated with a type I fimbria","authors":"Lesley Sitter, Marion Schoof, Travis R. Glare, Murray P. Cox, Peter C. Fineran, Paul P. Gardner, Mark R. H. Hurst","doi":"10.1002/mbo3.1395","DOIUrl":"10.1002/mbo3.1395","url":null,"abstract":"<p>A soil bacterium in the <i>Serratia</i> genus, carrying a 153 kb conjugative amber disease-associated plasmid (pADAP), is commercially exploited for population control of the New Zealand endemic pest beetle <i>Costelytra giveni</i> (Coleoptera: Scarabaeidae). The main insecticidal elements are an anti-feeding prophage and the Sep ABC toxin complex (Tc). Homologs of pADAP, encoding variant Tcs, convey different beetle disease phenotypes. To investigate the correlation between variable bioactivity and the Tc variant, 76 <i>Serratia</i> plasmids were sequenced, resulting in the identification of four additional <i>tc</i> variants. All <i>Serratia tc</i> variants were found to be colocated with a conserved type 1 <i>sef</i> fimbrial-like operon, indicating a conserved <i>sef-tc</i> genetic island not observed outside of the <i>Serratia</i> genus. The conserved co-location of the fimbrial and <i>tc</i> genes suggests the fimbriae somehow contribute to the lifestyle of Tc-producing cells. Expression of the <i>sef</i> operon in a fim-null <i>Escherichia coli</i> strain revealed fimbriae presence while a constructed <i>sef</i>-deficient mutant showed no reduction of virulence or host colonization. Although no detectable contribution of Sef to amber disease in <i>C. giveni</i> was observed, the Sef adhesin sequences clustered similarly to the <i>Serratia</i> species encoding it, suggesting Sef has a species-specific function.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 1","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1395","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139094476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Isini Buthgamuwa, Jane C. Fenelon, Alice Roser, Haley Meer, Stephen D. Johnston, Ashley M. Dungan
{"title":"Gut microbiota in the short-beaked echidna (Tachyglossus Aculeatus) shows stability across gestation","authors":"Isini Buthgamuwa, Jane C. Fenelon, Alice Roser, Haley Meer, Stephen D. Johnston, Ashley M. Dungan","doi":"10.1002/mbo3.1392","DOIUrl":"https://doi.org/10.1002/mbo3.1392","url":null,"abstract":"<p>Indigenous gut microbial communities (microbiota) play critical roles in health and may be especially important for the mother and fetus during pregnancy. Monotremes, such as the short-beaked echidna, have evolved to lay and incubate an egg, which hatches in their pouch where the young feeds. Since both feces and eggs pass through the cloaca, the fecal microbiota of female echidnas provides an opportunity for vertical transmission of microbes to their offspring. Here, we characterize the gut/fecal microbiome of female short-beaked echidnas and gain a better understanding of the changes that may occur in their microbiome as they go through pregnancy. Fecal samples from four female and five male echidnas were obtained from the Currumbin Wildlife Sanctuary in Queensland and sequenced to evaluate bacterial community structure. We identified 25 core bacteria, most of which were present in male and female samples. Genera such as <i>Fusobacterium</i>, <i>Bacteroides</i>, <i>Escherichia</i>-<i>Shigella</i>, and <i>Lactobacillus</i> were consistently abundant, regardless of sex or gestation stage, accounting for 58.00% and 56.14% of reads in male and female samples, respectively. The echidna microbiome remained stable across the different gestation stages, though there was a significant difference in microbiota composition between male and female echidnas. This study is the first to describe the microbiome composition of short-beaked echidnas across reproductive phases and allows the opportunity for this novel information to be used as a metric of health to aid in the detection of diseases triggered by microbiota dysbiosis.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 6","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1392","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138634348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Examining the functional space of gut microbiome-derived peptides","authors":"Ying-Chiang J. Lee","doi":"10.1002/mbo3.1393","DOIUrl":"https://doi.org/10.1002/mbo3.1393","url":null,"abstract":"<p>The human gut microbiome contains thousands of small, novel peptides that could play a role in microbe–microbe and host–microbe interactions, contributing to human health and disease. Although these peptides have not yet been systematically characterized, computational tools can be used to elucidate the bioactivities they may have. This article proposes probing the functional space of gut microbiome-derived peptides (MDPs) using in silico approaches for three bioactivities: antimicrobial, anticancer, and nucleomodulins. Machine learning programs that support peptide and protein queries are provided for each bioactivity. Considering the biases of an activity-centric approach, activity-agnostic tools using structural and chemical similarity and target prediction are also described. Gut MDPs represent a vast functional space that can not only contribute to our understanding of microbiome interactions but potentially even serve as a source of life-changing therapeutics.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 6","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1393","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138570903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Richard Wormald, Paul N. Humphreys, Christopher J. Charles, Simon P. Rout
{"title":"Bacillus-based probiotic cleansers reduce the formation of dry biofilms on common hospital surfaces","authors":"Richard Wormald, Paul N. Humphreys, Christopher J. Charles, Simon P. Rout","doi":"10.1002/mbo3.1391","DOIUrl":"https://doi.org/10.1002/mbo3.1391","url":null,"abstract":"<p>In the absence of liquid suspension, dry biofilms can form upon hard surfaces within a hospital environment, representing a healthcare-associated infection risk. Probiotic cleansers using generally recognized as safe organisms, such as those of the <i>Bacillus</i> genus, represent a potential strategy for the reduction of dry biofilm bioburden. The mechanisms of action and efficacy of these cleaners are, however, poorly understood. To address this, a preventative dry biofilm assay was developed using steel, melamine, and ceramic surfaces to assess the ability of a commercially available <i>Bacillus</i> spp. based probiotic cleanser to reduce the surface bioburden of <i>Escherichia coli</i> and <i>Staphylococcus aureus</i>. Via this assay, phosphate-buffered saline controls were able to generate dry biofilms within 7 days of incubation, with the application of the probiotic cleanser able to prevent >97.7% of dry biofilm formation across both pathogen analogs and surface types. Further to this, surfaces treated with the probiotic mixture alone also showed a reduction in dry biofilm across both pathogen and surface types. Confocal laser scanning microscopy imaging indicated that the probiotic bacteria were able to germinate and colonize surfaces, likely forming a protective layer upon these hard surfaces.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 6","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1391","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138431981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Correction to Concomitant knockout of target and transporter genes in filamentous fungi by genome coediting","authors":"","doi":"10.1002/mbo3.1390","DOIUrl":"10.1002/mbo3.1390","url":null,"abstract":"<p>Tamano, K. (2022). Concomitant knockout of target and transporter genes in filamentous fungi by genome coediting. MicrobiologyOpen, e1280. https://doi.org/10.1002/mbo3.1280</p><p>In the above article, the words “autotrophy” and “autotrophic” should be read as “prototrophy” and “prototrophic,” respectively in all instances.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 6","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1390","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135392869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Comparative genomic and metabolomic study of three Streptomyces sp. differing in biological activity","authors":"Alisson Gillon, Ola Abdelrahman, Eliane Abou-Mansour, Floriane L'Haridon, Laurent Falquet, Pierre-Marie Allard, Laure Weisskopf","doi":"10.1002/mbo3.1389","DOIUrl":"10.1002/mbo3.1389","url":null,"abstract":"<p>The <i>Streptomyces</i> genus is known to produce many specialized metabolites of value for medicine, but the potential of these metabolites in agronomy remains largely unexplored. In this study, we investigated three phylogenetically closely related <i>Streptomyces</i> strains (B5, B91, and B135) isolated from three distinct soil samples in Sudan. Despite belonging to the same species, these strains exhibited different ranges of <i>Phytophthora infestans</i> inhibition. The objective of this work was to identify the active compound(s) responsible for the inhibition of <i>P. infestans</i> and of other plant pathogens by comparing the genomes and metabolomes of the three strains which showed distinct activity patterns: B5 was the strongest inhibitor of oomycetes, B5 and B91 both inhibited most fungi and B135 was the only strain showing antibacterial activity. Our comparative genomic and metabolomic analysis identified borrelidin as the bioactive compound underlying B5's strong anti-oomycete activity and highlighted a few other metabolites as putative candidates underlying the strains' antifungal and antibacterial activities. This study illustrates the power of comparative genomics and metabolomics on phylogenetically closely related strains of differing activities to highlight bioactive compounds that could contribute to new sustainable crop protection strategies.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 6","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1389","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136069536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nadim Ahmad, Manfred Ritz, Anjuli Calchera, Jürgen Otte, Imke Schmitt, Thomas Brueck, Norbert Mehlmer
{"title":"Biosynthetic gene cluster synteny: Orthologous polyketide synthases in Hypogymnia physodes, Hypogymnia tubulosa, and Parmelia sulcata","authors":"Nadim Ahmad, Manfred Ritz, Anjuli Calchera, Jürgen Otte, Imke Schmitt, Thomas Brueck, Norbert Mehlmer","doi":"10.1002/mbo3.1386","DOIUrl":"https://doi.org/10.1002/mbo3.1386","url":null,"abstract":"<p>Lichens are symbiotic associations consisting of a photobiont (algae or cyanobacteria) and a mycobiont (fungus), which together generate a variety of unique secondary metabolites. To access this biosynthetic potential for biotechnological applications, deeper insights into the biosynthetic pathways and corresponding gene clusters are necessary. Here, we provide a comparative view of the biosynthetic gene clusters of three lichen mycobionts derived from <i>Hypogymnia physodes, Hypogymnia tubulosa</i>, and <i>Parmelia sulcata</i>. In addition, we present a high-quality PacBio metagenome of <i>Parmelia sulcata</i>, from which we extracted the mycobiont bin containing 214 biosynthetic gene clusters. Most biosynthetic gene clusters in these genomes were associated with T1PKSs, followed by NRPSs and terpenes. This study focused on biosynthetic gene clusters related to polyketide synthesis. Based on ketosynthase homology, we identified nine highly syntenic clusters present in all three species. Among the four clusters belonging to nonreducing PKSs, two are putatively linked to lichen substances derived from orsellinic acid (orcinol depsides and depsidones, e.g., lecanoric acid, physodic acid, lobaric acid), one to compounds derived from methylated forms of orsellinic acid (beta orcinol depsides, e.g., atranorin), and one to melanins. Five clusters with orthologs in all three species are linked to reducing PKSs. Our study contributes to sorting and dereplicating the vast PKS diversity found in lichenized fungi. High-quality sequences of biosynthetic gene clusters of these three common species provide a foundation for further exploration into biotechnological applications and the molecular evolution of lichen substances.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 5","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1386","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50136271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Helene R. Haugsten, Anne K. Kristoffersen, Trude M. Haug, Tine M. Søland, Reidun Øvstebø, Hans C. D. Aass, Morten Enersen, Hilde K. Galtung
{"title":"Isolation, characterization, and fibroblast uptake of bacterial extracellular vesicles from Porphyromonas gingivalis strains","authors":"Helene R. Haugsten, Anne K. Kristoffersen, Trude M. Haug, Tine M. Søland, Reidun Øvstebø, Hans C. D. Aass, Morten Enersen, Hilde K. Galtung","doi":"10.1002/mbo3.1388","DOIUrl":"https://doi.org/10.1002/mbo3.1388","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 <p>Periodontitis is an inflammatory condition caused by bacteria and represents a serious health problem worldwide as the inflammation damages the supporting tissues of the teeth and may predispose to systemic diseases. <i>Porphyromonas gingivalis</i> is considered a keystone periodontal pathogen that releases bacterial extracellular vesicles (bEVs) containing virulence factors, such as gingipains, that may contribute to the pathogenesis of periodontitis. This study aimed to isolate and characterize bEVs from three strains of <i>P. gingivalis</i>, investigate putative bEV uptake into human oral fibroblasts, and determine the gingipain activity of the bEVs. bEVs from three bacterial strains, ATCC 33277, A7A1-28, and W83, were isolated through ultrafiltration and size-exclusion chromatography. Vesicle size distribution was measured by nano-tracking analysis (NTA). Transmission electron microscopy was used for bEV visualization. Flow cytometry was used to detect bEVs and gingipain activity was measured with an enzyme assay using a substrate specific for arg-gingipain. The uptake of bEVs into oral fibroblasts was visualized using confocal microscopy. NTA showed bEV concentrations from 10<sup>8</sup> to 10<sup>11</sup> particles/mL and bEV diameters from 42 to 356 nm. TEM pictures demonstrated vesicle-like structures. bEV-gingipains were detected both by flow cytometry and enzyme assay. Fibroblasts incubated with bEVs labeled with fluorescent dye displayed intracellular localization consistent with bEV internalization. In conclusion, bEVs from <i>P. gingivalis</i> were successfully isolated and characterized, and their uptake into human oral fibroblasts was documented. The bEVs displayed active gingipains demonstrating their origin from <i>P. gingivalis</i> and the potential role of bEVs in periodontitis.</p>\u0000 </section>\u0000 </div>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 5","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1388","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50135494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}