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Transforming the untransformable with knockout minicircles: High-efficiency transformation and vector-free allelic exchange knockout in the fish pathogen Photobacterium damselae 用敲除微环转化不可转化物:鱼类病原体豆芽光杆菌的高效转化和无载体等位基因交换敲除。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-08-21 DOI: 10.1002/mbo3.1374
Oleksandra Rudenko, Laura Baseggio, Fynn McGuigan, Andrew C. Barnes
{"title":"Transforming the untransformable with knockout minicircles: High-efficiency transformation and vector-free allelic exchange knockout in the fish pathogen Photobacterium damselae","authors":"Oleksandra Rudenko,&nbsp;Laura Baseggio,&nbsp;Fynn McGuigan,&nbsp;Andrew C. Barnes","doi":"10.1002/mbo3.1374","DOIUrl":"10.1002/mbo3.1374","url":null,"abstract":"<p>Gene inactivation studies are critical in pathogenic bacteria, where insights into species biology can guide the development of vaccines and treatments. Allelic exchange via homologous recombination is a generic method of targeted gene editing in bacteria. However, generally applicable protocols are lacking, and suboptimal approaches are often used for nonstandard but epidemiologically important species. <i>Photobacterium damselae</i> subsp. <i>piscicida</i> (<i>Pdp</i>) is a primary pathogen of fish in aquaculture and has been considered hard to transform since the mid-1990s. Consequently, conjugative transfer of RK2/RP4 suicide vectors from <i>Escherichia coli</i> S17-1/SM10 donor strains, a system prone to off-target mutagenesis, was used to deliver the allelic exchange DNA in previous studies. Here we have achieved efficient electrotransformation in <i>Pdp</i> using a salt-free highly concentrated sucrose solution, which performs as a hypertonic wash buffer, cryoprotectant, and electroporation buffer. High-efficiency transformation has enabled vector-free mutagenesis for which we have employed circular minimalistic constructs (knockout minicircles) containing only allelic exchange essentials that were generated by Gibson assembly. Preparation of competent cells using sucrose and electroporation/integration of minicircles had virtually no detectable off-target promutagenic effect. In contrast, a downstream <i>sacB</i> selection apparently induced several large deletions via mobilization of transposable elements. Electroporation of minicircles into sucrose-treated cells is a versatile broadly applicable approach that may facilitate allelic exchange in a wide range of microbial species. The method permitted inactivation of a primary virulence factor unique to <i>Pdp</i>, apoptogenic toxin AIP56, demonstrating the efficacy of minicircles for difficult KO targets located on the high copy number of small plasmids.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141159","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}
引用次数: 0
Antibiofilm assay for antimicrobial peptides combating the sulfate-reducing bacteria Desulfovibrio vulgaris 抗硫酸盐还原菌寻常脱硫弧菌抗菌肽的抗体膜测定。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-08-21 DOI: 10.1002/mbo3.1376
Lena Stillger, Lucile Viau, Dirk Holtmann, Daniela Müller
{"title":"Antibiofilm assay for antimicrobial peptides combating the sulfate-reducing bacteria Desulfovibrio vulgaris","authors":"Lena Stillger,&nbsp;Lucile Viau,&nbsp;Dirk Holtmann,&nbsp;Daniela Müller","doi":"10.1002/mbo3.1376","DOIUrl":"10.1002/mbo3.1376","url":null,"abstract":"<p>In medical, environmental, and industrial processes, the accumulation of bacteria in biofilms can disrupt many processes. Antimicrobial peptides (AMPs) are receiving increasing attention in the development of new substances to avoid or reduce biofilm formation. There is a lack of parallel testing of the effect against biofilms in this area, as well as in the testing of other antibiofilm agents. In this paper, a high-throughput screening was developed for the analysis of the antibiofilm activity of AMPs, differentiated into inhibition and removal of a biofilm. The sulfate-reducing bacterium <i>Desulfovibrio vulgaris</i> was used as a model organism. <i>D. vulgaris</i> represents an undesirable bacterium, which is considered one of the major triggers of microbiologically influenced corrosion. The application of a 96-well plate and steel rivets as a growth surface realizes real-life conditions and at the same time establishes a flexible, simple, fast, and cost-effective assay. All peptides tested in this study demonstrated antibiofilm activity, although these peptides should be individually selected depending on the addressed aim. For biofilm inhibition, the peptide DASamP1 is the most suitable, with a sustained effect for up to 21 days. The preferred peptides for biofilm removal are S6L3-33, in regard to bacteria reduction, and Bactenecin, regarding total biomass reduction.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1376","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141160","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}
引用次数: 0
Cattle–compost–soil: The transfer of antibiotic resistance in livestock agriculture 牛堆肥土壤:畜牧农业中抗生素耐药性的转移。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-08-18 DOI: 10.1002/mbo3.1375
Fadhel Abbas, Phil Thomas, Bianca Cully-Duse, Nicholas M. Andronicos, Gal Winter
{"title":"Cattle–compost–soil: The transfer of antibiotic resistance in livestock agriculture","authors":"Fadhel Abbas,&nbsp;Phil Thomas,&nbsp;Bianca Cully-Duse,&nbsp;Nicholas M. Andronicos,&nbsp;Gal Winter","doi":"10.1002/mbo3.1375","DOIUrl":"10.1002/mbo3.1375","url":null,"abstract":"<p>Antibiotic resistance is a major global health threat. Agricultural use of antibiotics is considered to be a main contributor to the issue, influencing both animals and humans as defined by the One Health approach. The purpose of the present study was to determine the abundance of antibiotic-resistant bacterial populations and the overall bacterial diversity of cattle farm soils that have been treated with animal manure compost. Soil and manure samples were collected from different sites at Tullimba farm, NSW. Cultures were grown from these samples in the presence of 11 commonly used antibiotics and antibiotic-resistant bacteria (ARB) colonies were identified. Soil and manure bacterial diversity was also determined using 16S ribosomal RNA next-generation sequencing. Results showed that ARB abundance was greatest in fresh manure and significantly lower in composted manure. However, the application of composted manure on paddock soil led to a significant increase in soil ARB abundance. Of the antibiotics tested, the number of ARB in each sample was greatest for antibiotics that inhibited the bacterial cell wall and protein synthesis. Collectively, these results suggest that the transfer of antibiotic resistance from composted animal manure to soil may not be solely mediated through the application of live bacteria and highlight the need for further research into the mechanism of antibiotic resistance transfer.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1375","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141161","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}
引用次数: 0
Isolation and characterization of filamentous fungi capable of degrading the mycotoxin patulin 能够降解霉菌毒素棒曲霉素的丝状真菌的分离和鉴定。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-08-11 DOI: 10.1002/mbo3.1373
Megumi Mita, Rina Sato, Miho Kakinuma, Hiroyuki Nakagawa, Toshiki Furuya
{"title":"Isolation and characterization of filamentous fungi capable of degrading the mycotoxin patulin","authors":"Megumi Mita,&nbsp;Rina Sato,&nbsp;Miho Kakinuma,&nbsp;Hiroyuki Nakagawa,&nbsp;Toshiki Furuya","doi":"10.1002/mbo3.1373","DOIUrl":"10.1002/mbo3.1373","url":null,"abstract":"<p>Patulin is a toxic secondary metabolite synthesized by various fungal strains. This mycotoxin is generally toxic to microorganisms as well as mammals due to its reactivity with the important cellular antioxidant glutathione. In this study, we explored the presence of microorganisms capable of degrading patulin. Microorganisms were screened for the ability to both grow in culture medium containing patulin and reduce its concentration. Screening of 510 soil samples resulted in the isolation of two filamentous fungal strains, one of which, <i>Acremonium</i> sp. TUS-MM1 was characterized in detail. Liquid chromatography-mass spectrometry and nuclear magnetic resonance analyses revealed that TUS-MM1 cells degraded patulin to desoxypatulinic acid. In addition, extracellular components of strain TUS-MM1 also exhibited patulin-transforming activity. High-performance liquid chromatography analysis revealed that the extracellular components generated several products from patulin. Disc diffusion assay using <i>Escherichia coli</i> cells revealed that the patulin-transformation products by the extracellular components are less toxic than patulin. We also demonstrated that a thermostable, low-molecular-weight compound within the extracellular components was responsible for the patulin-transforming activity. These results suggest that strain TUS-MM1 transforms patulin into less-toxic molecules by secreting a highly reactive compound. In addition, once patulin enters the cells, strain TUS-MM1 can transform it into desoxypatulinic acid to reduce its toxicity.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1373","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141162","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}
引用次数: 0
Isolation, biochemical characterization, and genome sequencing of two high-quality genomes of a novel chitinolytic Jeongeupia species 一个新的溶壳Jeongeupia物种的两个高质量基因组的分离、生化表征和基因组测序。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-08-06 DOI: 10.1002/mbo3.1372
Nathanael D. Arnold, Daniel Garbe, Thomas B. Brück
{"title":"Isolation, biochemical characterization, and genome sequencing of two high-quality genomes of a novel chitinolytic Jeongeupia species","authors":"Nathanael D. Arnold,&nbsp;Daniel Garbe,&nbsp;Thomas B. Brück","doi":"10.1002/mbo3.1372","DOIUrl":"10.1002/mbo3.1372","url":null,"abstract":"<p>Chitin is the second most abundant polysaccharide worldwide as part of arthropods' exoskeletons and fungal cell walls. Low concentrations in soils and sediments indicate rapid decomposition through chitinolytic organisms in terrestrial and aquatic ecosystems. The enacting enzymes, so-called chitinases, and their products, chitooligosaccharides, exhibit promising characteristics with applications ranging from crop protection to cosmetics, medical, textile, and wastewater industries. Exploring novel chitinolytic organisms is crucial to expand the enzymatical toolkit for biotechnological chitin utilization and to deepen our understanding of diverse catalytic mechanisms. In this study, we present two long-read sequencing-based genomes of highly similar <i>Jeongeupia</i> species, which have been screened, isolated, and biochemically characterized from chitin-amended soil samples. Through metabolic characterization, whole-genome alignments, and phylogenetic analysis, we could demonstrate how the investigated strains differ from the taxonomically closest strain <i>Jeongeupia naejangsanensis</i> BIO-TAS4-2<sup>T</sup> (DSM 24253). In silico analysis and sequence alignment revealed a multitude of highly conserved chitinolytic enzymes in the investigated <i>Jeongeupia</i> genomes. Based on these results, we suggest that the two strains represent a novel species within the genus of <i>Jeongeupia</i>, which may be useful for environmentally friendly <i>N</i>-acetylglucosamine production from crustacean shell or fungal biomass waste or as a crop protection agent.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1372","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10149742","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}
引用次数: 1
Co-localization of clinically relevant antibiotic- and heavy metal resistance genes on plasmids in Klebsiella pneumoniae from marine bivalves 海洋双壳类肺炎克雷伯菌质粒上临床相关抗生素和重金属抗性基因的共定位。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-07-19 DOI: 10.1002/mbo3.1368
Fredrik Håkonsholm, Marit A. K. Hetland, Iren H. Löhr, Bjørn Tore Lunestad, Nachiket P. Marathe
{"title":"Co-localization of clinically relevant antibiotic- and heavy metal resistance genes on plasmids in Klebsiella pneumoniae from marine bivalves","authors":"Fredrik Håkonsholm,&nbsp;Marit A. K. Hetland,&nbsp;Iren H. Löhr,&nbsp;Bjørn Tore Lunestad,&nbsp;Nachiket P. Marathe","doi":"10.1002/mbo3.1368","DOIUrl":"10.1002/mbo3.1368","url":null,"abstract":"<p><i>Klebsiella pneumoniae</i> is an opportunistic pathogen frequently associated with antibiotic resistance and present in a wide range of environments, including marine habitats. However, little is known about the development, persistence, and spread of antibiotic resistance in such environments. This study aimed to obtain the complete genome sequences of antibiotic-resistant <i>K. pneumoniae</i> isolated from marine bivalves in order to determine the genetic context of antibiotic- and heavy metal resistance genes in these isolates. Five antibiotic-resistant <i>K. pneumoniae</i> isolates, of which four also carried heavy metal resistance genes, were selected for complete genome sequencing using the Illumina MiSeq platform and the Oxford Nanopore Technologies GridION device. Conjugation experiments were conducted to examine the transfer potential of selected plasmids. The average length of the complete genomes was 5.48 Mbp with a mean chromosome size of 5.27 Mbp. Seven plasmids were detected in the antibiotic-resistant isolates. Three IncFIB, one IncFIB/IncFII, and one IncFIB/IncHIB plasmid, respectively, carried antibiotic resistance genes such as <i>qnrS1, aph(6)-Id</i> and <i>aph(3′)-Ia, aadA1</i>, and <i>aadA2</i>. Four of these plasmids also carried genes encoding resistance to copper (<i>pco</i>), silver (<i>sil</i>), and arsenic (<i>ars</i>). One plasmid carrying <i>tet(D</i>) and <i>bla</i><sub>SHV-1</sub> as well as <i>pco, sil</i>, and <i>ars</i> genes was transferred to <i>Escherichia coli</i> by conjugation. We show the co-occurrence of antibiotic- and heavy metal resistance genes on a conjugative IncFIB plasmid from <i>K. pneumoniae</i> from marine bivalves. Our study highlights the importance of the marine environment and seafood as a possible dissemination route for antimicrobial resistance and provides insights into the potential for co-selection of antibiotic resistance genes by heavy metals.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1368","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141166","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}
引用次数: 0
Isolation and functional analysis of phage-displayed antibody fragments targeting the staphylococcal superantigen-like proteins 噬菌体展示的针对葡萄球菌超抗原样蛋白的抗体片段的分离和功能分析。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-07-16 DOI: 10.1002/mbo3.1371
Ida Alanko, Rebecca Sandberg, Eeva-Christine Brockmann, Carla J. C. de Haas, Jos A. G. van Strijp, Urpo Lamminmäki, Outi M. H. Salo-Ahen
{"title":"Isolation and functional analysis of phage-displayed antibody fragments targeting the staphylococcal superantigen-like proteins","authors":"Ida Alanko,&nbsp;Rebecca Sandberg,&nbsp;Eeva-Christine Brockmann,&nbsp;Carla J. C. de Haas,&nbsp;Jos A. G. van Strijp,&nbsp;Urpo Lamminmäki,&nbsp;Outi M. H. Salo-Ahen","doi":"10.1002/mbo3.1371","DOIUrl":"10.1002/mbo3.1371","url":null,"abstract":"<p><i>Staphylococcus aureus</i> produces numerous virulence factors that manipulate the immune system, helping the bacteria avoid phagocytosis. In this study, we are investigating three immune evasion molecules called the staphylococcal superantigen-like proteins 1, 5, and 10 (SSL1, SSL5, and SSL10). All three SSLs inhibit vital host immune processes and contribute to <i>S</i>. <i>aureus</i> immune evasion. This study aimed to identify single-chain variable fragment (scFvs) antibodies from synthetic antibody phage libraries, which can recognize either of the three SSLs and could block the interaction between the SSLs and their respective human targets. The antibodies were isolated after three rounds of panning against SSL1, SSL5, and SSL10, and their ability to bind to the SSLs was studied using a time-resolved fluorescence-based immunoassay. We successfully obtained altogether 44 unique clones displaying binding activity to either SSL1, SSL5, or SSL10. The capability of the SSL-recognizing scFvs to inhibit the SSLs' function was tested in an MMP9 enzymatic activity assay, a P-selectin glycoprotein ligand 1 competitive binding assay, and an IgG1-mediated phagocytosis assay. We could show that one scFv was able to inhibit SSL1 and maintain MMP9 activity in a concentration-dependent manner. Finally, the structure of this inhibiting scFv was modeled and used to create putative scFv-SSL1-complex models by protein–protein docking. The complex models were subjected to a 100-ns molecular dynamics simulation to assess the possible binding mode of the antibody.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1371","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10149741","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}
引用次数: 0
Clay-associated microbial communities and their relevance for a nuclear waste repository in the Opalinus Clay rock formation Opalinus粘土岩层中粘土相关微生物群落及其与核废料库的相关性。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-07-10 DOI: 10.1002/mbo3.1370
Julia Mitzscherling, Steffi Genderjahn, Anja M. Schleicher, Alexander Bartholomäus, Jens Kallmeyer, Dirk Wagner
{"title":"Clay-associated microbial communities and their relevance for a nuclear waste repository in the Opalinus Clay rock formation","authors":"Julia Mitzscherling,&nbsp;Steffi Genderjahn,&nbsp;Anja M. Schleicher,&nbsp;Alexander Bartholomäus,&nbsp;Jens Kallmeyer,&nbsp;Dirk Wagner","doi":"10.1002/mbo3.1370","DOIUrl":"10.1002/mbo3.1370","url":null,"abstract":"<p>Microorganisms are known to be natural agents of biocorrosion and mineral transformation, thereby potentially affecting the safety of deep geological repositories used for high-level nuclear waste storage. To better understand how resident microbial communities of the deep terrestrial biosphere may act on mineralogical and geochemical characteristics of insulating clays, we analyzed their structure and potential metabolic functions, as well as site-specific mineralogy and element composition from the dedicated Mont Terri underground research laboratory, Switzerland. We found that the Opalinus Clay formation is mainly colonized by Alphaproteobacteria, Firmicutes, and Bacteroidota, which are known for corrosive biofilm formation. Potential iron-reducing bacteria were predominant in comparison to methanogenic archaea and sulfate-reducing bacteria. Despite microbial communities in Opalinus Clay being in majority homogenous, site-specific mineralogy and geochemistry conditions have selected for subcommunities that display metabolic potential for mineral dissolution and transformation. Our findings indicate that the presence of a potentially low-active mineral-associated microbial community must be further studied to prevent effects on the repository's integrity over the long term.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 4","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1370","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10141165","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}
引用次数: 0
Morphological and physiological impacts of salinity on colonial strains of the cyanobacteria Microcystis aeruginosa 盐度对铜绿微囊藻蓝藻群落菌株形态和生理的影响
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-06-28 DOI: 10.1002/mbo3.1367
Myriam Bormans, Benjamin Legrand, Nicolas Waisbord, Enora Briand
{"title":"Morphological and physiological impacts of salinity on colonial strains of the cyanobacteria Microcystis aeruginosa","authors":"Myriam Bormans,&nbsp;Benjamin Legrand,&nbsp;Nicolas Waisbord,&nbsp;Enora Briand","doi":"10.1002/mbo3.1367","DOIUrl":"10.1002/mbo3.1367","url":null,"abstract":"<p>In the context of global change and enhanced toxic cyanobacterial blooms, cyanobacterial transfer to estuaries is likely to increase in frequency and intensity and impact animal and human health. Therefore, it is important to evaluate the potential of their survival in estuaries. In particular, we tested if the colonial form generally observed in natural blooms enhanced the resistance to salinity shock compared to the unicellular form generally observed in isolated strains. We tested the impact of salinity on two colonial strains of <i>Microcystis aeruginosa</i>, producing different amounts of mucilage by combining classical batch methods with a novel microplate approach. We demonstrate that the collective organization of these pluricellular colonies improves their ability to cope with osmotic shock when compared to unicellular strains. The effect of a sudden high salinity increase (<i>S</i> ≥ 20) over 5 to 6 days had several impacts on the morphology of <i>M. aeruginosa</i> colonies. For both strains, we observed a gradual increase in colony size and a gradual decrease in intercellular spacing. For one strain, we also observed a decrease in cell diameter with an increase in mucilage extent. The pluricellular colonies formed by both strains could withstand higher salinities than unicellular strains studied previously. In particular, the strain producing more mucilage displayed a sustained autofluorescence even at <i>S</i> = 20, a limit that is higher than the most robust unicellular strain. These results imply survival and possible <i>M. aeruginosa</i> proliferation in mesohaline estuaries.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1367","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9731858","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}
引用次数: 2
An improved method for intracellular DNA (iDNA) recovery from terrestrial environments 从陆地环境中提取细胞内DNA (iDNA)的改进方法
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2023-06-25 DOI: 10.1002/mbo3.1369
Diego Medina Caro, Lucas Horstmann, Lars Ganzert, Romulo Oses, Thomas Friedl, Dirk Wagner
{"title":"An improved method for intracellular DNA (iDNA) recovery from terrestrial environments","authors":"Diego Medina Caro,&nbsp;Lucas Horstmann,&nbsp;Lars Ganzert,&nbsp;Romulo Oses,&nbsp;Thomas Friedl,&nbsp;Dirk Wagner","doi":"10.1002/mbo3.1369","DOIUrl":"10.1002/mbo3.1369","url":null,"abstract":"<p>The simultaneous extraction of intracellular DNA (iDNA) and extracellular DNA (eDNA) can help to separate the living in situ community (represented by iDNA) from background DNA that originated both from past communities and from allochthonous sources. As iDNA and eDNA extraction protocols require separating cells from the sample matrix, their DNA yields are generally lower than direct methods that lyse the cells within the sample matrix. We, therefore, tested different buffers with and without adding a detergent mix (DM) in the extraction protocol to improve the recovery of iDNA from surface and subsurface samples that covered a variety of terrestrial environments. The combination of a highly concentrated sodium phosphate buffer plus DM significantly improved iDNA recovery for almost all tested samples. Additionally, the combination of sodium phosphate and EDTA improved iDNA recovery in most of the samples and even allowed the successful extraction of iDNA from extremely low-biomass iron-bearing rock samples taken from the deep biosphere. Based on our results, we recommend using a protocol with sodium phosphate in combination with either a DM (NaP 300 mM + DM) or EDTA (NaP + EDTA 300 mM). Furthermore, for studies that rely on the eDNA pool, we recommend using buffers solely based on sodium phosphate because the addition of EDTA or a DM resulted in a decrease in eDNA for most of the tested samples. These improvements can help reduce community bias in environmental studies and contribute to better characterizations of both modern and past ecosystems.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"12 3","pages":""},"PeriodicalIF":3.4,"publicationDate":"2023-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1369","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9726448","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}
引用次数: 0
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