-
Cureus Oct 2020is an uncommon pathogen in humans and there are no documented cases of infection associated with central venous catheters. Here we describe a 72-year-old man on...
is an uncommon pathogen in humans and there are no documented cases of infection associated with central venous catheters. Here we describe a 72-year-old man on hemodialysis who presented with a fever and was found to have bacteremia. The only obvious source of infection was the hemodialysis catheter. The isolate was susceptible to all antibiotics tested and he was successfully treated with ciprofloxacin and central venous catheter removal. Patients with chronic medical conditions and vascular devices are at risk for invasive infections with rare Pseudomonas species. As laboratory pathogen detection advances arise, it is possible that additional cases of infections in humans will be identified. Our case provides one example of the successful treatment of bacteremia in a 72-year-old man with a line-associated infection.
PubMed: 33178507
DOI: 10.7759/cureus.10853 -
Microbiology Spectrum Jun 2023Several variants of the plasmid-carried tigecycline resistance gene cluster, , have been identified. This study characterized another novel variant, , located on the...
Several variants of the plasmid-carried tigecycline resistance gene cluster, , have been identified. This study characterized another novel variant, , located on the chromosome of environmental-origin Pseudomonas mendocina. TMexC6D6-TOprJ1 mediates resistance to multiple drugs, including tigecycline. The promoter activity of and negative transcriptional repression by the upstream regulator tnfxB6 are crucial for the expression of . was found in the plasmids or chromosomes of different Pseudomonas species from six countries. Two genetic backgrounds, class 1 integrons and -carrying integrase units, were found adjacent to the gene cluster and might mediate the transfer of this novel efflux pump gene cluster in Pseudomonas. Further phylogenetic analysis revealed Pseudomonas as the major reservoir of variants, warranting closer monitoring in the future. Tigecycline is one of the treatment options for serious infections caused by multidrug-resistant bacteria, and tigecycline resistance has gained extensive attention. The emergence of a transferable tigecycline resistance efflux pump gene cluster, , severely challenged the efficiency of tigecycline. In this study, we identified another novel variant, , which could confer resistance to multiple classes of antibiotics, including tigecycline. Although was found only in Pseudomonas species, might spread to hosts via mobile genetic elements resembling those of other variants, compromising the therapeutic strategies. Meanwhile, novel transferable variants are constantly emerging and mostly exist in Pseudomonas spp., indicating Pseudomonas as the important hidden reservoir and origin of variants. Continuous monitoring and investigations of are urgent to control its spread.
Topics: Tigecycline; Pseudomonas; Phylogeny; Anti-Bacterial Agents; Plasmids; Microbial Sensitivity Tests
PubMed: 37067462
DOI: 10.1128/spectrum.00767-23 -
BioRxiv : the Preprint Server For... Oct 2023Previously, we discovered that a small RNA from a clinical isolate of PA14, induces learned avoidance and its transgenerational inheritance in . is an important human...
Previously, we discovered that a small RNA from a clinical isolate of PA14, induces learned avoidance and its transgenerational inheritance in . is an important human pathogen, and there are other in natural habitat, but it is unclear whether ever encounters PA14-like bacteria in the wild. Thus, it is not known if small RNAs from bacteria found in natural habitat can also regulate host behavior and produce heritable behavioral effects. Here we found that a pathogenic strain isolated from the microbiota, GRb0427, like PA14, regulates worm behavior: worms learn to avoid this pathogenic bacterium following exposure to GRb0427, and this learned avoidance is inherited for four generations. The learned response is entirely mediated by bacterially-produced small RNAs, which induce avoidance and transgenerational inheritance, providing further support that such mechanisms of learning and inheritance exist in the wild. Using bacterial small RNA sequencing, we identified Pv1, a small RNA from GRb0427, that matches the sequence of . We find that Pv1 is both necessary and sufficient to induce learned avoidance of Grb0427. However, Pv1 also results in avoidance of a beneficial microbiome strain, ; this potentially maladaptive response may favor reversal of the transgenerational memory after a few generations. Our findings suggest that bacterial small RNA-mediated regulation of host behavior and its transgenerational inheritance are functional in natural environment, and that different bacterial small RNA-mediated regulation systems evolved independently but define shared molecular features of bacterial small RNAs that produce transgenerationally-inherited effects.
PubMed: 37503135
DOI: 10.1101/2023.07.20.549962 -
Applied and Environmental Microbiology May 2021The widely prescribed pharmaceutical metformin and its main metabolite, guanylurea, are currently two of the most common contaminants in surface and wastewater....
The widely prescribed pharmaceutical metformin and its main metabolite, guanylurea, are currently two of the most common contaminants in surface and wastewater. Guanylurea often accumulates and is poorly, if at all, biodegraded in wastewater treatment plants. This study describes strain GU, isolated from a municipal wastewater treatment plant, using guanylurea as its sole nitrogen source. The genome was sequenced with 36-fold coverage and mined to identify guanylurea degradation genes. The gene encoding the enzyme initiating guanylurea metabolism was expressed, and the enzyme was purified and characterized. Guanylurea hydrolase, a newly described enzyme, was shown to transform guanylurea to one equivalent (each) of ammonia and guanidine. Guanidine also supports growth as a sole nitrogen source. Cell yields from growth on limiting concentrations of guanylurea revealed that metabolism releases all four nitrogen atoms. Genes encoding complete metabolic transformation were identified bioinformatically, defining the pathway as follows: guanylurea to guanidine to carboxyguanidine to allophanate to ammonia and carbon dioxide. The first enzyme, guanylurea hydrolase, is a member of the isochorismatase-like hydrolase protein family, which includes biuret hydrolase and triuret hydrolase. Although homologs, the three enzymes show distinct substrate specificities. Pairwise sequence comparisons and the use of sequence similarity networks allowed fine structure discrimination between the three homologous enzymes and provided insights into the evolutionary origins of guanylurea hydrolase. Metformin is a pharmaceutical most prescribed for type 2 diabetes and is now being examined for potential benefits to COVID-19 patients. People taking the drug pass it largely unchanged, and it subsequently enters wastewater treatment plants. Metformin has been known to be metabolized to guanylurea. The levels of guanylurea often exceed that of metformin, leading to the former being considered a "dead-end" metabolite. Metformin and guanylurea are water pollutants of emerging concern, as they persist to reach nontarget aquatic life and humans, the latter if it remains in treated water. The present study has identified a strain that completely degrades guanylurea. The genome was sequenced, and the genes involved in guanylurea metabolism were identified in three widely separated genomic regions. This knowledge advances the idea that guanylurea is not a dead-end product and will allow for bioinformatic identification of the relevant genes in wastewater treatment plant microbiomes and other environments subjected to metagenomic sequencing.
Topics: Ammonia; Bacterial Proteins; Biodegradation, Environmental; Biomineralization; Genome, Bacterial; Guanidine; Hydrolases; Metabolic Networks and Pathways; Metformin; Multigene Family; Pseudomonas mendocina; Substrate Specificity; Urea; Wastewater; Water Pollutants, Chemical
PubMed: 33741630
DOI: 10.1128/AEM.03003-20 -
Scientific Reports Dec 2022PmlR2, a class II LitR/CarH family transcriptional regulator, and PmSB-LOV, a "short" LOV-type blue light photoreceptor, are adjacently encoded in Pseudomonas mendocina...
PmlR2, a class II LitR/CarH family transcriptional regulator, and PmSB-LOV, a "short" LOV-type blue light photoreceptor, are adjacently encoded in Pseudomonas mendocina NBRC 14162. An effector protein for the "short" LOV-type photoreceptor in Pseudomonas has not yet been identified. Here, we show that PmlR2 is an effector protein of PmSB-LOV. Transcriptional analyses revealed that the expression of genes located near pmlR2 and its homolog gene, pmlR1, was induced in response to illumination. In vitro DNA-protein binding analyses showed that recombinant PmlR2 directly binds to the promoter region of light-inducible genes. Furthermore PmSB-LOV exhibited a typical LOV-type light-induced spectral change. Gel-filtration chromatography demonstrated that the illuminated PmSB-LOV was directly associated with PmlR2, whereas non-illuminated proteins did not interact. The inhibition of PmlR2 function following PmSB-LOV binding was verified by surface plasmon resonance: the DNA-binding ability of PmlR2 was specifically inhibited in the presence of blue light-illuminated-PmSB-LOV. An In vitro transcription assay showed a dose-dependent reduction in PmlR2 repressor activity in the presence of illuminated PmSB-LOV. Overall, evidence suggests that the DNA-binding activity of PmlR2 is inhibited by its direct association with blue light-activated PmSB-LOV, enabling transcription of light-inducible promoters by RNA polymerase.
Topics: Pseudomonas mendocina; Bacterial Proteins; Promoter Regions, Genetic; Protein Binding; DNA
PubMed: 36526696
DOI: 10.1038/s41598-022-26254-3 -
3 Biotech Sep 2019This study aimed to investigate the effects of cytoskeleton protein MreB on bacterial cell morphology and the synthesis of alginate oligosaccharides (AO) and...
This study aimed to investigate the effects of cytoskeleton protein MreB on bacterial cell morphology and the synthesis of alginate oligosaccharides (AO) and polyhydroxyalkanoate (PHA) by NK-01. To overexpress the gene, an expression vector encoding MreB-GFP fusion protein was constructed. The scanning electron microscope (SEM) showed that cells expressing MreB were longer than the wild ones, which agrees with MreB's relationship with the synthesis of peptidoglycan. Cells expressing the MreB-GFP fusion protein emitted green fluorescence under a fluorescence microscope, suggesting that MreB was functionally expressed in strain NK-01. Under a confocal laser scanning microscope, MreB was observed as located around the cell membrane. Furthermore, the recombinant strain could synthesize 0.961 g/L AO, which was 5.86-fold higher than wild-type strain. Through the medium optimization test, we finally selected the addition of 20 g/L glucose as the optimal glycogen addition for AO fermentation based on a high AO yield and high substrate transformation efficiency. The results indicated that overexpression of MreB affected the cell morphology, the activity of AO polymerase, and the efficiency of AO secretion. However, the synthesis of PHA for recombinant strain was slightly reduced. The results suggested that the overexpression of this cytoskeleton protein affected the yield of specific intracellular and extracellular products.
PubMed: 31497462
DOI: 10.1007/s13205-019-1873-7 -
Foods (Basel, Switzerland) Mar 2023Previous research has shown that freshwater edible fish imported into Australia are not compliant with Australian importation guidelines and as a result may be high risk...
Previous research has shown that freshwater edible fish imported into Australia are not compliant with Australian importation guidelines and as a result may be high risk for bacterial contamination. In the present study, the outer surface of imported freshwater fish were swabbed, cultured, confirmatory tests performed and antimicrobial patterns investigated. Channidae fish (Sp. A/n = 66) were contaminated with zoonotic sp./ (n = 1/66) and other bacteria implicated in cases of opportunistic human infection, these being sp. (including . and (n = 34/66)); sp. (n = 32/66); (n = 27/66) and (n = 3/66). Pangasiidae fish (Species B/n = 47) were contaminated with zoonotic (n = 10/47); sp. (n = 6/47) and environmental bacteria sp. (n = 3/47). One sample was resistant to all antimicrobials tested and is considered to be Methicillin Resistant . Mud, natural diet, or vegetation identified in Sp. A fish/or packaging were significantly associated with the presence of spp. The study also showed that visibly clean fish (Sp. B) may harbour zoonotic bacteria and that certain types of bacteria are common to fish groups, preparations, and contaminants. Further investigations are required to support the development of appropriate food safety recommendations in Australia.
PubMed: 36981215
DOI: 10.3390/foods12061288 -
PLoS Genetics Mar 2024C. elegans can learn to avoid pathogenic bacteria through several mechanisms, including bacterial small RNA-induced learned avoidance behavior, which can be inherited...
C. elegans can learn to avoid pathogenic bacteria through several mechanisms, including bacterial small RNA-induced learned avoidance behavior, which can be inherited transgenerationally. Previously, we discovered that a small RNA from a clinical isolate of Pseudomonas aeruginosa, PA14, induces learned avoidance and transgenerational inheritance of that avoidance in C. elegans. Pseudomonas aeruginosa is an important human pathogen, and there are other Pseudomonads in C. elegans' natural habitat, but it is unclear whether C. elegans ever encounters PA14-like bacteria in the wild. Thus, it is not known if small RNAs from bacteria found in C. elegans' natural habitat can also regulate host behavior and produce heritable behavioral effects. Here we screened a set of wild habitat bacteria, and found that a pathogenic Pseudomonas vranovensis strain isolated from the C. elegans microbiota, GRb0427, regulates worm behavior: worms learn to avoid this pathogenic bacterium following exposure, and this learned avoidance is inherited for four generations. The learned response is entirely mediated by bacterially-produced small RNAs, which induce avoidance and transgenerational inheritance, providing further support that such mechanisms of learning and inheritance exist in the wild. We identified Pv1, a small RNA expressed in P. vranovensis, that has a 16-nucleotide match to an exon of the C. elegans gene maco-1. Pv1 is both necessary and sufficient to induce learned avoidance of Grb0427. However, Pv1 also results in avoidance of a beneficial microbiome strain, P. mendocina. Our findings suggest that bacterial small RNA-mediated regulation of host behavior and its transgenerational inheritance may be functional in C. elegans' natural environment, and that this potentially maladaptive response may favor reversal of the transgenerational memory after a few generations. Our data also suggest that different bacterial small RNA-mediated regulation systems evolved independently, but define shared molecular features of bacterial small RNAs that produce transgenerationally-inherited effects.
Topics: Animals; Humans; Caenorhabditis elegans; Caenorhabditis elegans Proteins; RNA, Small Interfering; RNA Interference; RNA, Bacterial; Pseudomonas aeruginosa; Bacteria
PubMed: 38547071
DOI: 10.1371/journal.pgen.1011178 -
Foods (Basel, Switzerland) Nov 2022Alginate lyases (ALyases) have been widely applied in enzymatically degrading alginate for the preparation of alginate oligosaccharides (AOS), which possess a range of...
Alginate lyases (ALyases) have been widely applied in enzymatically degrading alginate for the preparation of alginate oligosaccharides (AOS), which possess a range of excellent physiological benefits including immunoregulatory, antivirus, and antidiabetic properties. Among the characterized ALyases, the number of ALyases with strict substrate specificity which possess potential in directed preparation of AOS is quite small. ALyases of polysaccharides lyase (PL) 5 family have been reported to perform poly--D-mannuronic acid (Poly-M) substrate specificity. However, there have been fewer studies with a comprehensive characterization and comparison of PL 5 family ALyases. In this study, a putative PL 5 family ALyase PMD was cloned from and expressed in . The novel ALyase presented maximum activity at 30 °C and pH 7.0. PMD displayed pH stability properties under the range of pH 5 to pH 9, which retained more than 80% relative activity, even when incubated for 48 h. Product analysis indicated that PMD might be an endolytic ALyase with strict Poly M substrate specificity and yield disaccharide and trisaccharide as main products. In addition, residues K58, R66, Y248, and R344 were proposed to be the potential key residues for catalysis via site-directed mutation. Detailed characterization of PMD and comprehensive comparisons could supply some different information about properties of PL 5 ALyases which might be helpful for its application in the directed production of AOS.
PubMed: 36360141
DOI: 10.3390/foods11213527 -
MSystems Apr 2024Plant-associated diazotrophs strongly relate to plant nitrogen (N) supply and growth. However, our knowledge of diazotrophic community assembly and microbial N...
Plant-associated diazotrophs strongly relate to plant nitrogen (N) supply and growth. However, our knowledge of diazotrophic community assembly and microbial N metabolism in plant microbiomes is largely limited. Here we examined the assembly and temporal dynamics of diazotrophic communities across multiple compartments (soils, epiphytic and endophytic niches of root and leaf, and grain) of three cereal crops (maize, wheat, and barley) and identified the potential N-cycling pathways in phylloplane microbiomes. Our results demonstrated that the microbial species pool, influenced by site-specific environmental factors (e.g., edaphic factors), had a stronger effect than host selection (i.e., plant species and developmental stage) in shaping diazotrophic communities across the soil-plant continuum. Crop diazotrophic communities were dominated by a few taxa (~0.7% of diazotrophic phylotypes) which were mainly affiliated with , , , and . Furthermore, eight dominant taxa belonging to and were identified as keystone diazotrophic taxa for three crops and were potentially associated with microbial network stability and crop yields. Metagenomic binning recovered 58 metagenome-assembled genomes (MAGs) from the phylloplane, and the majority of them were identified as novel species (37 MAGs) and harbored genes potentially related to multiple N metabolism processes (e.g., nitrate reduction). Notably, for the first time, a high-quality MAG harboring genes involved in the complete denitrification process was recovered in the phylloplane and showed high identity to . Overall, these findings significantly expand our understanding of ecological drivers of crop diazotrophs and provide new insights into the potential microbial N metabolism in the phyllosphere.IMPORTANCEPlants harbor diverse nitrogen-fixing microorganisms (i.e., diazotrophic communities) in both belowground and aboveground tissues, which play a vital role in plant nitrogen supply and growth promotion. Understanding the assembly and temporal dynamics of crop diazotrophic communities is a prerequisite for harnessing them to promote plant growth. In this study, we show that the site-specific microbial species pool largely shapes the structure of diazotrophic communities in the leaves and roots of three cereal crops. We further identify keystone diazotrophic taxa in crop microbiomes and characterize potential microbial N metabolism pathways in the phyllosphere, which provides essential information for developing microbiome-based tools in future sustainable agricultural production.
Topics: Microbiota; Agriculture; Soil; Nitrogen; Crops, Agricultural; Plant Development
PubMed: 38501864
DOI: 10.1128/msystems.01055-23