Robyn Dove, Emily R. Wolfe, Nathan U. Stewart, Abigail Coleman, Sara Herrejon Chavez, Daniel J. Ballhorn
The root nodules of actinorhizal plants are home to nitrogen-fixing bacterial symbionts, known as Frankia, along with a small percentage of other microorganisms. These include fungal endophytes and non-Frankia bacteria. The taxonomic and functional diversity of the microbial consortia within these root nodules is not well understood. In this study, we surveyed and analyzed the cultivable, non-Frankia fungal and bacterial endophytes of root nodules from red and Sitka alder trees that grow together. We examined their taxonomic diversity, co-occurrence, differences between hosts, and potential functional roles. For the first time, we are reporting numerous fungal endophytes of alder root nodules. These include Sporothrix guttuliformis, Fontanospora sp., Cadophora melinii, an unclassified Cadophora, Ilyonectria destructans, an unclassified Gibberella, Nectria ramulariae, an unclassified Trichoderma, Mycosphaerella tassiana, an unclassified Talaromyces, Coniochaeta sp., and Sistotrema brinkmanii. We are also reporting several bacterial genera for the first time: Collimonas, Psychrobacillus, and Phyllobacterium. Additionally, we are reporting the genus Serratia for the second time, with the first report having been recently published in 2023. Pseudomonas was the most frequently isolated bacterial genus and was found to co-inhabit individual nodules with both fungi and bacteria. We found that the communities of fungal endophytes differed by host species, while the communities of bacterial endophytes did not.
{"title":"Root nodules of red alder (Alnus rubra) and sitka alder (Alnus viridis ssp. sinuata) are inhabited by taxonomically diverse cultivable microbial endophytes","authors":"Robyn Dove, Emily R. Wolfe, Nathan U. Stewart, Abigail Coleman, Sara Herrejon Chavez, Daniel J. Ballhorn","doi":"10.1002/mbo3.1422","DOIUrl":"10.1002/mbo3.1422","url":null,"abstract":"<p>The root nodules of actinorhizal plants are home to nitrogen-fixing bacterial symbionts, known as <i>Frankia</i>, along with a small percentage of other microorganisms. These include fungal endophytes and non-<i>Frankia</i> bacteria. The taxonomic and functional diversity of the microbial consortia within these root nodules is not well understood. In this study, we surveyed and analyzed the cultivable, non-<i>Frankia</i> fungal and bacterial endophytes of root nodules from red and Sitka alder trees that grow together. We examined their taxonomic diversity, co-occurrence, differences between hosts, and potential functional roles. For the first time, we are reporting numerous fungal endophytes of alder root nodules. These include <i>Sporothrix guttuliformis</i>, <i>Fontanospora</i> sp., <i>Cadophora melinii</i>, an unclassified <i>Cadophora</i>, <i>Ilyonectria destructans</i>, an unclassified <i>Gibberella</i>, <i>Nectria ramulariae</i>, an unclassified <i>Trichoderma</i>, <i>Mycosphaerella tassiana</i>, an unclassified <i>Talaromyces</i>, <i>Coniochaeta</i> sp., and <i>Sistotrema brinkmanii</i>. We are also reporting several bacterial genera for the first time: <i>Collimonas</i>, <i>Psychrobacillus</i>, and <i>Phyllobacterium</i>. Additionally, we are reporting the genus <i>Serratia</i> for the second time, with the first report having been recently published in 2023. <i>Pseudomonas</i> was the most frequently isolated bacterial genus and was found to co-inhabit individual nodules with both fungi and bacteria. We found that the communities of fungal endophytes differed by host species, while the communities of bacterial endophytes did not.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1422","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141284230","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}
Daniel P. R. Herlemann, Helen Tammert, Carmen Kivistik, Kairi Käiro, Veljo Kisand
The factors that influence the distribution of bacterial community composition are not well understood. The role of geographical patterns, which suggest limited dispersal, is still a topic of debate. Bacteria associated with hosts face unique dispersal challenges as they often rely on their hosts, which provide specific environments for their symbionts. In this study, we examined the effect of biogeographic distances on the bacterial diversity and composition of bacterial communities in the gastrointestinal tract of Ampullaceana balthica. We compared the effects on the host-associated bacterial community to those on bacterial communities in water and sediment. This comparison was made using 16S ribosomal RNA gene sequencing. We found that the bacterial communities we sampled in Estonia, Denmark, and Northern Germany varied between water, sediment, and the gastrointestinal tract. They also varied between countries within each substrate. This indicates that the type of substrate is a dominant factor in determining bacterial community composition. We separately analyzed the turnover rates of water, sediment, and gastrointestinal bacterial communities over increasing geographic distances. We observed that the turnover rate was lower for gastrointestinal bacterial communities compared to water bacterial communities. This implies that the composition of gastrointestinal bacteria remains relatively stable over distances, while water bacterial communities exhibit greater variability. However, the gastrointestinal tract had the lowest percentage of country-specific amplicon sequence variants, suggesting bacterial colonization from local bacterial communities. Since the overlap between the water and gastrointestinal tract was highest, it appears that the gastrointestinal bacterial community is colonized by the water bacterial community. Our study confirmed that biogeographical patterns in host-associated communities differ from those in water and sediment bacterial communities. These host-associated communities consist of numerous facultative symbionts derived from the water bacterial community.
{"title":"Distinct biogeographical patterns in snail gastrointestinal tract bacterial communities compared with sediment and water","authors":"Daniel P. R. Herlemann, Helen Tammert, Carmen Kivistik, Kairi Käiro, Veljo Kisand","doi":"10.1002/mbo3.1413","DOIUrl":"10.1002/mbo3.1413","url":null,"abstract":"<p>The factors that influence the distribution of bacterial community composition are not well understood. The role of geographical patterns, which suggest limited dispersal, is still a topic of debate. Bacteria associated with hosts face unique dispersal challenges as they often rely on their hosts, which provide specific environments for their symbionts. In this study, we examined the effect of biogeographic distances on the bacterial diversity and composition of bacterial communities in the gastrointestinal tract of <i>Ampullaceana balthica</i>. We compared the effects on the host-associated bacterial community to those on bacterial communities in water and sediment. This comparison was made using 16S ribosomal RNA gene sequencing. We found that the bacterial communities we sampled in Estonia, Denmark, and Northern Germany varied between water, sediment, and the gastrointestinal tract. They also varied between countries within each substrate. This indicates that the type of substrate is a dominant factor in determining bacterial community composition. We separately analyzed the turnover rates of water, sediment, and gastrointestinal bacterial communities over increasing geographic distances. We observed that the turnover rate was lower for gastrointestinal bacterial communities compared to water bacterial communities. This implies that the composition of gastrointestinal bacteria remains relatively stable over distances, while water bacterial communities exhibit greater variability. However, the gastrointestinal tract had the lowest percentage of country-specific amplicon sequence variants, suggesting bacterial colonization from local bacterial communities. Since the overlap between the water and gastrointestinal tract was highest, it appears that the gastrointestinal bacterial community is colonized by the water bacterial community. Our study confirmed that biogeographical patterns in host-associated communities differ from those in water and sediment bacterial communities. These host-associated communities consist of numerous facultative symbionts derived from the water bacterial community.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1413","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141200308","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}
The standard method of receptor activation involves the binding of signals or signal-loaded solute binding proteins (SBPs) to sensor domains. Many sensor histidine kinases (SHKs), which are activated by SBP binding, are encoded adjacent to their corresponding sbp gene. We examined three SBPs of Pseudomonas aeruginosa PAO1, encoded near the genes for the AgtS (PA0600) and AruS (PA4982) SHKs, to determine how common this arrangement is. Ligand screening and microcalorimetric studies revealed that the SBPs PA0602 and PA4985 preferentially bind to GABA (KD = 2.3 and 0.58 μM, respectively), followed by 5-aminovalerate (KD = 30 and 1.6 μM, respectively) and ethanoldiamine (KD = 2.3 and 0.58 μM, respectively). In contrast, AgtB (PA0604) exclusively recognizes 5-aminovaleric acid (KD = 2.9 μM). However, microcalorimetric titrations did not show any binding between the AgtS sensor domain and AgtB or PA0602, regardless of the presence of ligands. Similarly, bacterial two-hybrid assays did not demonstrate an interaction between PA4985 and the AruS sensor domain. Therefore, sbp and shk genes located nearby are not always functionally linked. We previously identified PA0222 as a GABA-specific SBP. The presence of three SBPs for GABA may be linked to GABA's role as a trigger for P. aeruginosa virulence.
{"title":"Exploring solute binding proteins in Pseudomonas aeruginosa that bind to γ-aminobutyrate and 5-aminovalerate and their role in activating sensor kinases","authors":"Jean Paul Cerna-Vargas, Tino Krell","doi":"10.1002/mbo3.1415","DOIUrl":"10.1002/mbo3.1415","url":null,"abstract":"<p>The standard method of receptor activation involves the binding of signals or signal-loaded solute binding proteins (SBPs) to sensor domains. Many sensor histidine kinases (SHKs), which are activated by SBP binding, are encoded adjacent to their corresponding sbp gene. We examined three SBPs of <i>Pseudomonas aeruginosa</i> PAO1, encoded near the genes for the AgtS (PA0600) and AruS (PA4982) SHKs, to determine how common this arrangement is. Ligand screening and microcalorimetric studies revealed that the SBPs PA0602 and PA4985 preferentially bind to GABA (KD = 2.3 and 0.58 μM, respectively), followed by 5-aminovalerate (KD = 30 and 1.6 μM, respectively) and ethanoldiamine (KD = 2.3 and 0.58 μM, respectively). In contrast, AgtB (PA0604) exclusively recognizes 5-aminovaleric acid (KD = 2.9 μM). However, microcalorimetric titrations did not show any binding between the AgtS sensor domain and AgtB or PA0602, regardless of the presence of ligands. Similarly, bacterial two-hybrid assays did not demonstrate an interaction between PA4985 and the AruS sensor domain. Therefore, sbp and shk genes located nearby are not always functionally linked. We previously identified PA0222 as a GABA-specific SBP. The presence of three SBPs for GABA may be linked to GABA's role as a trigger for <i>P. aeruginosa</i> virulence.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1415","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141081786","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}
Indrawattana N., Pumipuntu N., Suriyakhun N., Jangsangthong A., Kulpeanprasit S., Chantratita N., Sookrung N., Chaicumpa W., & Buranasinsup S. Staphylococcus argenteus from rabbits in Thailand. MicrobiologyOpen. 2018;e665.
In “abstract” section, line 4, the text “the presence of S. argenteus in isolated S. aureus was investigated in 67 rabbits with abscess lesions during 2014–2016.” should read as “the presence of S. argenteus in isolated S. aureus was investigated in 67 rabbits with abscess lesions during 2014–2015.”
In “material and method section, 2.1 | Specimen collection and bacterial isolation, line 3, the text “Sixty-seven pus samples were collected from rabbits with clinical abscesses by a veterinarian at Prasu-Arthorn Animal Hospital, Thailand during 2014–2016.” should read as “Sixty-seven pus samples were collected from rabbits with clinical abscesses by a veterinarian at Prasu-Arthorn Animal Hospital, Thailand, during 2014–2015.”
We apologize for this error.
Indrawattana N., Pumipuntu N., Suriyakhun N., Jangsangthong A., Kulpeanprasit S., Chantratita N., Sookrung N., Chaicumpa W., & Buranasinsup S. 来自泰国兔子的金黄色葡萄球菌。MicrobiologyOpen.2018;e665.In "abstract" section, line 4, the text "the presence of S. argenteus in isolated S. aureus was investigated in 67 rabbits with abscess lesions during 2014-2016." should read as "the presence of S. argenteus in isolated S. aureus was investigated in 67 rabbits with abscess lesions during 2014-2015. "In "material and method section, 2.1 | 标本采集和细菌分离 "中,第 3 行,"2014-2016 年期间,泰国 Prasu-Arthorn 动物医院的一名兽医从患有临床脓肿的兔子身上采集了 67 份脓液样本 "应为 "2014-2015 年期间,泰国 Prasu-Arthorn 动物医院的一名兽医从患有临床脓肿的兔子身上采集了 67 份脓液样本"。
{"title":"Correction to “Staphylococcus argenteus from rabbits in Thailand”","authors":"","doi":"10.1002/mbo3.1414","DOIUrl":"10.1002/mbo3.1414","url":null,"abstract":"<p>Indrawattana N., Pumipuntu N., Suriyakhun N., Jangsangthong A., Kulpeanprasit S., Chantratita N., Sookrung N., Chaicumpa W., & <b>Buranasinsup S</b>. <i>Staphylococcus argenteus</i> from rabbits in Thailand. <i>MicrobiologyOpen</i>. 2018;e665.</p><p>In “abstract” section, line 4, the text “the presence of <i>S. argenteus</i> in isolated <i>S. aureus</i> was investigated in 67 rabbits with abscess lesions during 2014–2016.” should read as “the presence of <i>S. argenteus</i> in isolated <i>S. aureus</i> was investigated in 67 rabbits with abscess lesions during 2014–2015.”</p><p>In “material and method section, 2.1 | Specimen collection and bacterial isolation, line 3, the text “Sixty-seven pus samples were collected from rabbits with clinical abscesses by a veterinarian at Prasu-Arthorn Animal Hospital, Thailand during 2014–2016.” should read as “Sixty-seven pus samples were collected from rabbits with clinical abscesses by a veterinarian at Prasu-Arthorn Animal Hospital, Thailand, during 2014–2015.”</p><p>We apologize for this error.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1414","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140922647","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}
Judith Stiefelmaier, Joshua Keller, Wiebke Neupert, Roland Ulber
Cable bacteria, characterized by their multicellular filamentous growth, are prevalent in both freshwater and marine sediments. They possess the unique ability to transport electrons over distances of centimeters. Coupled with their capacity to fix CO2 and their record-breaking conductivity for biological materials, these bacteria present promising prospects for bioprocess engineering, including potential electrochemical applications. However, the cultivation of cable bacteria has been limited to their natural sediment, constraining their utility in production processes. To address this, our study designs synthetic sediment, drawing on ion exchange chromatography data from natural sediments and existing literature on the requirements of cable bacteria. We examined the effects of varying bentonite concentrations on water retention and the impacts of different sands. For the first time, we cultivated cable bacteria on synthetic sediment, specifically the freshwater strain Electronema aureum GS. This cultivation was conducted over 10 weeks in a specially developed sediment bioreactor, resulting in an increased density of cable bacteria in the sediment and growth up to a depth of 5 cm. The creation of this synthetic sediment paves the way for the reproducible cultivation of cable bacteria. It also opens up possibilities for future process scale-up using readily available components. This advancement holds significant implications for the broader field of bioprocess engineering.
{"title":"Towards bioprocess engineering of cable bacteria: Establishment of a synthetic sediment","authors":"Judith Stiefelmaier, Joshua Keller, Wiebke Neupert, Roland Ulber","doi":"10.1002/mbo3.1412","DOIUrl":"https://doi.org/10.1002/mbo3.1412","url":null,"abstract":"<p>Cable bacteria, characterized by their multicellular filamentous growth, are prevalent in both freshwater and marine sediments. They possess the unique ability to transport electrons over distances of centimeters. Coupled with their capacity to fix CO<sub>2</sub> and their record-breaking conductivity for biological materials, these bacteria present promising prospects for bioprocess engineering, including potential electrochemical applications. However, the cultivation of cable bacteria has been limited to their natural sediment, constraining their utility in production processes. To address this, our study designs synthetic sediment, drawing on ion exchange chromatography data from natural sediments and existing literature on the requirements of cable bacteria. We examined the effects of varying bentonite concentrations on water retention and the impacts of different sands. For the first time, we cultivated cable bacteria on synthetic sediment, specifically the freshwater strain <i>Electronema aureum</i> GS. This cultivation was conducted over 10 weeks in a specially developed sediment bioreactor, resulting in an increased density of cable bacteria in the sediment and growth up to a depth of 5 cm. The creation of this synthetic sediment paves the way for the reproducible cultivation of cable bacteria. It also opens up possibilities for future process scale-up using readily available components. This advancement holds significant implications for the broader field of bioprocess engineering.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1412","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140844664","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}
Ciara Kenneally, Craig P. Murphy, Roy D. Sleator, Eamonn P. Culligan
Traditional bacteriocin screening methods often face limitations due to diffusion-related challenges in agar matrices, which can prevent the peptides from reaching their target organism. Turbidimetric techniques offer a solution to these issues, eliminating diffusion-related problems and providing an initial quantification of bacteriocin efficacy in producer organisms. This study involved screening the cell-free supernatant (CFS) from eight uncharacterized asymptomatic bacteriuria (ABU) isolates and Escherichia coli 83972 for antimicrobial activity against clinical uropathogenic E. coli (UPEC) strains using turbidimetric growth methods. ABU isolates exhibiting activity against five or more UPEC strains were further characterized (PUTS 37, PUTS 58, PUTS 59, S-07-4, and SK-106-1). The inhibition of the CFS by proteinase K suggested that the antimicrobial activity was proteinaceous in nature, potentially bacteriocins. The activity of E. coli PUTS 58 and SK-106-1 was enhanced in an artificial urine medium, with both inhibiting all eight UPECs. A putative microcin H47 operon was identified in E. coli SK-106-1, along with a previously identified microcin V and colicin E7 in E. coli PUTS 37 and PUTS 58, respectively. These findings indicate that ABU bacteriocin-producers could serve as viable prophylactics and therapeutics in the face of increasing antibiotic resistance among uropathogens.
{"title":"Turbidimetric bioassays: A solution to antimicrobial activity detection in asymptomatic bacteriuria isolates against uropathogenic Escherichia coli","authors":"Ciara Kenneally, Craig P. Murphy, Roy D. Sleator, Eamonn P. Culligan","doi":"10.1002/mbo3.1411","DOIUrl":"https://doi.org/10.1002/mbo3.1411","url":null,"abstract":"<p>Traditional bacteriocin screening methods often face limitations due to diffusion-related challenges in agar matrices, which can prevent the peptides from reaching their target organism. Turbidimetric techniques offer a solution to these issues, eliminating diffusion-related problems and providing an initial quantification of bacteriocin efficacy in producer organisms. This study involved screening the cell-free supernatant (CFS) from eight uncharacterized asymptomatic bacteriuria (ABU) isolates and <i>Escherichia coli</i> 83972 for antimicrobial activity against clinical uropathogenic <i>E. coli</i> (UPEC) strains using turbidimetric growth methods. ABU isolates exhibiting activity against five or more UPEC strains were further characterized (PUTS 37, PUTS 58, PUTS 59, S-07-4, and SK-106-1). The inhibition of the CFS by proteinase K suggested that the antimicrobial activity was proteinaceous in nature, potentially bacteriocins. The activity of <i>E. coli</i> PUTS 58 and SK-106-1 was enhanced in an artificial urine medium, with both inhibiting all eight UPECs. A putative microcin H47 operon was identified in <i>E. coli</i> SK-106-1, along with a previously identified microcin V and colicin E7 in <i>E. coli</i> PUTS 37 and PUTS 58, respectively. These findings indicate that ABU bacteriocin-producers could serve as viable prophylactics and therapeutics in the face of increasing antibiotic resistance among uropathogens.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1411","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140844765","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}
Faizan Saleem, Enze Li, Kevin L. Tran, Bashudev Rudra, Thomas A. Edge, Herb E. Schellhorn, Radhey S. Gupta
Escherichia coli serves as a proxy indicator of fecal contamination in aquatic ecosystems. However, its identification using traditional culturing methods can take up to 24 h. The application of DNA markers, such as conserved signature proteins (CSPs) genes (unique to all species/strains of a specific taxon), can form the foundation for novel polymerase chain reaction (PCR) tests that unambiguously identify and detect targeted bacterial taxa of interest. This paper reports the identification of three new highly-conserved CSPs (genes), namely YahL, YdjO, and YjfZ, which are exclusive to E. coli/Shigella. Using PCR primers based on highly conserved regions within these CSPs, we have developed quantitative PCR (qPCR) assays for the evaluation of E. coli/Shigella species in water ecosystems. Both in-silico and experimental PCR testing confirmed the absence of sequence match when tested against other bacteria, thereby confirming 100% specificity of the tested CSPs for E. coli/Shigella. The qPCR assays for each of the three CSPs provided reliable quantification for all tested enterohaemorrhagic and environmental E. coli strains, a requirement for water testing. For recreational water samples, CSP-based quantification showed a high correlation (r > 7, p < 0.01) with conventional viable E. coli enumeration. This indicates that novel CSP-based qPCR assays for E. coli can serve as robust tools for monitoring water ecosystems and other critical areas, including food monitoring.
{"title":"Utilizing novel Escherichia coli-specific conserved signature proteins for enhanced monitoring of recreational water quality","authors":"Faizan Saleem, Enze Li, Kevin L. Tran, Bashudev Rudra, Thomas A. Edge, Herb E. Schellhorn, Radhey S. Gupta","doi":"10.1002/mbo3.1410","DOIUrl":"https://doi.org/10.1002/mbo3.1410","url":null,"abstract":"<p><i>Escherichia coli</i> serves as a proxy indicator of fecal contamination in aquatic ecosystems. However, its identification using traditional culturing methods can take up to 24 h. The application of DNA markers, such as conserved signature proteins (CSPs) genes (unique to all species/strains of a specific taxon), can form the foundation for novel polymerase chain reaction (PCR) tests that unambiguously identify and detect targeted bacterial taxa of interest. This paper reports the identification of three new highly-conserved CSPs (genes), namely <i>YahL</i>, <i>YdjO</i>, and <i>YjfZ</i>, which are exclusive to <i>E. coli</i>/<i>Shigella</i>. Using PCR primers based on highly conserved regions within these CSPs, we have developed quantitative PCR (qPCR) assays for the evaluation of <i>E. coli</i>/<i>Shigella</i> species in water ecosystems. Both in-silico and experimental PCR testing confirmed the absence of sequence match when tested against other bacteria, thereby confirming 100% specificity of the tested CSPs for <i>E. coli</i>/<i>Shigella</i>. The qPCR assays for each of the three CSPs provided reliable quantification for all tested enterohaemorrhagic and environmental <i>E. coli</i> strains, a requirement for water testing. For recreational water samples, CSP-based quantification showed a high correlation (<i>r</i> > 7, <i>p</i> < 0.01) with conventional viable <i>E. coli</i> enumeration. This indicates that novel CSP-based qPCR assays for <i>E. coli</i> can serve as robust tools for monitoring water ecosystems and other critical areas, including food monitoring.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1410","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140808160","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}
Sylvia A. Sapula, Bradley J. Hart, Naomi L. Siderius, Anteneh Amsalu, Jack M. Blaikie, Henrietta Venter
Stenotrophomonas maltophilia is a multidrug-resistant (MDR), Gram-negative bacterium intrinsically resistant to beta-lactams, including last-resort carbapenems. As an opportunistic pathogen, it can cause serious healthcare-related infections. This study assesses the prevalence, resistance profiles, and genetic diversity of S. maltophilia isolated from residential aged care facilities (RACFs). RACFs are known for their overuse and often inappropriate use of antibiotics, creating a strong selective environment that favors the development of bacterial resistance. The study was conducted on 73 S. maltophilia isolates recovered from wastewater and facility swab samples obtained from three RACFs and a retirement village. Phenotypic and genotypic assessments of the isolates revealed high carbapenem resistance, exemplifying their intrinsic beta-lactam resistance. Alarmingly, 49.3% (36/73) of the isolates were non-wild type for colistin, with minimum inhibitory concentration values of > 4 mg/L, and 11.0% (8/73) were resistant to trimethoprim-sulfamethoxazole. No resistance mechanisms were detected for either antimicrobial. Genotypic assessment of known lineages revealed isolates clustering with Sm17 and Sm18, lineages not previously reported in Australia, suggesting the potential ongoing spread of MDR S. maltophilia. Lastly, although only a few isolates were biocide tolerant (2.7%, 2/73), their ability to grow in high concentrations (64 mg/L) of triclosan is concerning, as it may be selecting for their survival and continued dissemination.
{"title":"Multidrug-resistant Stenotrophomonas maltophilia in residential aged care facilities: An emerging threat","authors":"Sylvia A. Sapula, Bradley J. Hart, Naomi L. Siderius, Anteneh Amsalu, Jack M. Blaikie, Henrietta Venter","doi":"10.1002/mbo3.1409","DOIUrl":"https://doi.org/10.1002/mbo3.1409","url":null,"abstract":"<p><i>Stenotrophomonas maltophilia</i> is a multidrug-resistant (MDR), Gram-negative bacterium intrinsically resistant to beta-lactams, including last-resort carbapenems. As an opportunistic pathogen, it can cause serious healthcare-related infections. This study assesses the prevalence, resistance profiles, and genetic diversity of <i>S. maltophilia</i> isolated from residential aged care facilities (RACFs). RACFs are known for their overuse and often inappropriate use of antibiotics, creating a strong selective environment that favors the development of bacterial resistance. The study was conducted on 73 <i>S. maltophilia</i> isolates recovered from wastewater and facility swab samples obtained from three RACFs and a retirement village. Phenotypic and genotypic assessments of the isolates revealed high carbapenem resistance, exemplifying their intrinsic beta-lactam resistance. Alarmingly, 49.3% (36/73) of the isolates were non-wild type for colistin, with minimum inhibitory concentration values of > 4 mg/L, and 11.0% (8/73) were resistant to trimethoprim-sulfamethoxazole. No resistance mechanisms were detected for either antimicrobial. Genotypic assessment of known lineages revealed isolates clustering with Sm17 and Sm18, lineages not previously reported in Australia, suggesting the potential ongoing spread of MDR <i>S. maltophilia</i>. Lastly, although only a few isolates were biocide tolerant (2.7%, 2/73), their ability to grow in high concentrations (64 mg/L) of triclosan is concerning, as it may be selecting for their survival and continued dissemination.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1409","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140808144","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}
Microbial communities from various environments have been studied in the quest for new natural products with a broad range of applications in medicine and biotechnology. We employed an enrichment method and genome mining tools to examine the biosynthetic potential of microbial communities in the sediments of a coastal sinkhole within the karst ecosystem of the Yucatán Peninsula, Mexico. Our investigation led to the detection of 203 biosynthetic gene clusters (BGCs) and 55 secondary metabolites (SMs) within 35 high-quality metagenome-assembled genomes (MAGs) derived from these subcommunities. The most abundant types of BGCs were Terpene, Nonribosomal peptide-synthetase, and Type III polyketide synthase. Some of the in silico identified BGCs and SMs have been previously reported to exhibit biological activities against pathogenic bacteria and fungi. Others could play significant roles in the sinkhole ecosystem, such as iron solubilization and osmotic stress protection. Interestingly, 75% of the BGCs showed no sequence homology with bacterial BGCs previously reported in the MiBIG database. This suggests that the microbial communities in this environment could be an untapped source of genes encoding novel specialized compounds. The majority of the BGCs were identified in pathways found in the genus Virgibacillus, followed by Sporosarcina, Siminovitchia, Rhodococcus, and Halomonas. The latter, along with Paraclostridium and Lysinibacillus, had the highest number of identified BGC types. This study offers fresh insights into the potential ecological role of SMs from sediment microbial communities in an unexplored environment, underscoring their value as a source of novel natural products.
{"title":"Biosynthetic potential of the sediment microbial subcommunities of an unexplored karst ecosystem and its ecological implications","authors":"Pablo Suárez-Moo, Alejandra Prieto-Davó","doi":"10.1002/mbo3.1407","DOIUrl":"https://doi.org/10.1002/mbo3.1407","url":null,"abstract":"<p>Microbial communities from various environments have been studied in the quest for new natural products with a broad range of applications in medicine and biotechnology. We employed an enrichment method and genome mining tools to examine the biosynthetic potential of microbial communities in the sediments of a coastal sinkhole within the karst ecosystem of the Yucatán Peninsula, Mexico. Our investigation led to the detection of 203 biosynthetic gene clusters (BGCs) and 55 secondary metabolites (SMs) within 35 high-quality metagenome-assembled genomes (MAGs) derived from these subcommunities. The most abundant types of BGCs were Terpene, Nonribosomal peptide-synthetase, and Type III polyketide synthase. Some of the in silico identified BGCs and SMs have been previously reported to exhibit biological activities against pathogenic bacteria and fungi. Others could play significant roles in the sinkhole ecosystem, such as iron solubilization and osmotic stress protection. Interestingly, 75% of the BGCs showed no sequence homology with bacterial BGCs previously reported in the MiBIG database. This suggests that the microbial communities in this environment could be an untapped source of genes encoding novel specialized compounds. The majority of the BGCs were identified in pathways found in the genus <i>Virgibacillus</i>, followed by <i>Sporosarcina, Siminovitchia, Rhodococcus</i>, and <i>Halomonas</i>. The latter, along with <i>Paraclostridium</i> and <i>Lysinibacillus</i>, had the highest number of identified BGC types. This study offers fresh insights into the potential ecological role of SMs from sediment microbial communities in an unexplored environment, underscoring their value as a source of novel natural products.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 2","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1407","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140541138","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}
Ian M. Lillie, Charles E. Booth, Adelaide E. Horvath, Matthew Mondragon, Melinda A. Engevik, Thomas D. Horvath
Arginine-ornithine metabolism plays a crucial role in bacterial homeostasis, as evidenced by numerous studies. However, the utilization of arginine and the downstream products of its metabolism remain undefined in various gut bacteria. To bridge this knowledge gap, we employed genomic screening to pinpoint relevant metabolic targets. We also devised a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics method to measure the levels of arginine, its upstream precursors, and downstream products in cell-free conditioned media from enteric pathobionts, including Escherichia coli, Klebsiella aerogenes, K. pneumoniae, Pseudomonas fluorescens, Acinetobacter baumannii, Streptococcus agalactiae, Staphylococcus epidermidis, S. aureus, and Enterococcus faecalis. Our findings revealed that all selected bacterial strains consumed glutamine, glutamate, and arginine, and produced citrulline, ornithine, and GABA in our chemically defined medium. Additionally, E. coli, K. pneumoniae, K. aerogenes, and P. fluorescens were found to convert arginine to agmatine and produce putrescine. Interestingly, arginine supplementation promoted biofilm formation in K. pneumoniae, while ornithine supplementation enhanced biofilm formation in S. epidermidis. These findings offer a comprehensive insight into arginine-ornithine metabolism in enteric pathobionts.
{"title":"Characterizing arginine, ornithine, and putrescine pathways in enteric pathobionts","authors":"Ian M. Lillie, Charles E. Booth, Adelaide E. Horvath, Matthew Mondragon, Melinda A. Engevik, Thomas D. Horvath","doi":"10.1002/mbo3.1408","DOIUrl":"https://doi.org/10.1002/mbo3.1408","url":null,"abstract":"<p>Arginine-ornithine metabolism plays a crucial role in bacterial homeostasis, as evidenced by numerous studies. However, the utilization of arginine and the downstream products of its metabolism remain undefined in various gut bacteria. To bridge this knowledge gap, we employed genomic screening to pinpoint relevant metabolic targets. We also devised a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics method to measure the levels of arginine, its upstream precursors, and downstream products in cell-free conditioned media from enteric pathobionts, including <i>Escherichia coli</i>, <i>Klebsiella aerogenes</i>, <i>K. pneumoniae</i>, <i>Pseudomonas fluorescens</i>, <i>Acinetobacter baumannii</i>, <i>Streptococcus agalactiae</i>, <i>Staphylococcus epidermidis</i>, <i>S. aureus</i>, and <i>Enterococcus faecalis</i>. Our findings revealed that all selected bacterial strains consumed glutamine, glutamate, and arginine, and produced citrulline, ornithine, and GABA in our chemically defined medium. Additionally, <i>E. coli</i>, <i>K. pneumoniae</i>, <i>K. aerogenes</i>, and <i>P. fluorescens</i> were found to convert arginine to agmatine and produce putrescine. Interestingly, arginine supplementation promoted biofilm formation in <i>K. pneumoniae</i>, while ornithine supplementation enhanced biofilm formation in <i>S. epidermidis</i>. These findings offer a comprehensive insight into arginine-ornithine metabolism in enteric pathobionts.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 2","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1408","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140333096","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}