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Lactobacillus plantarum is a potential starter and health-promoting probiotic bacterium. Effective, precise and diverse genome-editing of Lactobacillus plantarum without introducing exogenous genes or plasmids is of great importance. In this study, CRISPR/Cas9-assisted dsDNA and ssDNA recombineering were established in L. plantarum WCFS1 to seamlessly edit the genome, including gene knockout, insertions and point mutations. To optimize our editing method, phosphorothioate modification was used to improve the dsDNA insertion, and adenine-specific methyltransferase was used to improve the ssDNA recombination efficiency. These strategies were applied to engineer L. plantarum WCFS1 towards producing N-acetylglucosamine (GlcNAc). nagB was trunca.
This study reports that a high concentration of the endo-β-1,3-glucanase ENG (200 μg mL -1 ) induced heat-inactivated stipe wall extension of C. cinerea , whereas a high concentration of the extracellular β-glucosidase BGL2 (1000 μg mL -1 ) did not; however, in combination, low concentrations of ENG (25 μg mL -1 ) and BGL2 (260 μg mL -1 ) induced heat-inactivated stipe cell wall extension.
Production of unconventional oil and gas continues to rise, but the effects of high-density hydraulic fracturing (HF) activity near aquatic ecosystems are not fully understood. A commonly used biocide in HF, 2,2-dibromo-3-nitrilopropionamide (DBNPA), was studied in microcosms of HF-impacted vs. HF-unimpacted surface water streams to (1) compare the microbial community response, (2) investigate DBNPA degradation products based on past HF exposure, and (3) compare the microbial community response differences and similarities between the HF biocides DBNPA and glutaraldehyde. The microbial community responded to DBNPA differently in HF-impacted vs. HF-unimpacted microcosms in terms of 16S rRNA gene copies quantified, alpha and beta diversity, a.
Bovine respiratory disease (BRD) is a major cause of morbidity and mortality in beef cattle. Recent evidence suggests that commensal bacteria of the bovine nasopharynx have an important role in maintaining respiratory health by providing colonization resistance against pathogens. The objective of this study was to screen and select bacterial therapeutic candidates from the nasopharynx of feedlot cattle to mitigate the BRD pathogen Mannheimia haemolytica . In a stepwise approach, bacteria (n = 300) isolated from the nasopharynx of 100 healthy feedlot cattle were identified and initially screened (n = 178 isolates from 12 different genera) for growth inhibition of M. haemolytica . Subsequently, selected isolates were evaluated for their abili.
Cyclic diguanosine monophosphate (c-di-GMP) is an important second messenger involved in bacterial switching from motile to sessile lifestyles. In the opportunistic pathogen Pseudomonas aeruginosa , at least 40 genes are predicted to encode proteins for the making and breaking of this signal molecule. However, there is still paucity of information concerning the systemic expression pattern of those genes and the functions of uncharacterized genes. In this study, we analyzed the phylogenetic distribution of genes from P. aeruginosa that predicted to have a GGDEF domain and found five genes (PA5487, PA0285, PA0290, PA4367 and PA5017) with highly conserved distribution across 52 public complete Pseudomonads genomes. PA5487 was further characte.
Microbially influenced corrosion (MIC) results in significant damage to metallic materials in many industries. Anaerobic sulfate reducing bacteria (SRB) have been well-studied for their involvement in these processes. Highly corrosive environments are also found in pulp and paper processing where chloride and thiosulfate lead to corrosion of stainless steels. Acidithiobacillus ferrooxidans are critically important chemolithotrophic acidophiles exploited in metal biomining operations and there is interest in using these cells for emerging processes such as electronic waste recycling. We explored conditions where A. ferrooxidans could enable corrosion of stainless steel. Acidic media with iron, chloride, low sulfate and pyrite supplementation.
DDT (dichloro-diphenyl-trichloroethane) was used worldwide as an organochlorine insecticide to control agricultural pests and vectors of several insect-borne human diseases. It was banned in most industrialized countries, however, due to its persistence in the environment, DDT residues remain in environmental compartments becoming long-term sources of exposure. To identify and select fungal species suitable for bioremediation of DDT-contaminated sites, soil samples were collected from DDT-contaminated agricultural soils in Poland and thirty eight fungal taxa among 18 genera were isolated. Two of them, Trichoderma hamatum FBL 587 and Rhizopus arrhizus FBL 578, were tested for tolerance in the presence of 1 mg l -1 DDT concentration by using
Culture-based molecular identification methods have revolutionized detection of pathogens, yet these methods are slow and may yield inconclusive results from environmental materials. The second-generation sequencing tools have much improved precision and sensitivity of detection, but these analyses are costly and may take several days to months. Of the third-generation sequencing techniques, the portable MinION device (Oxford Nanopore Technologies) has received much attention because of its small size and possibility of rapid analysis at reasonable cost. Here, we compare the relative performance of two third-generation sequencing instruments, MinION and Sequel (Pacific Biosciences) in identification and diagnostics of fungal and oomycete pa.
Yeasts form mutualistic interactions with insects. Hallmarks of this interaction include provision of essential nutrients, while insects facilitate yeast dispersal and growth on plant substrates. A phylogenetically ancient, chemical dialogue coordinates this interaction, where the vocabulary, the volatile chemicals that mediate the insect response, remains largely unknown. Here, we employed gas chromatography-mass spectrometry (GC-MS), followed by hierarchical cluster (HCA) and orthogonal partial least square discriminant analysis (OPLS-DA), to profile the volatomes of six Metschnikowia spp., Cryptococcus nemorosus and brewer's yeast Saccharomyces cerevisiae . The yeasts, which are all found in association with insects feeding on foliage or.
Thioredoxin (Trx) and glutaredoxin (Grx) antioxidant systems are deeply involved in bacterial response to oxidative stress, but to date we know surprisingly little about roles of these systems in response to reactive oxygen species (ROS) other than hydrogen peroxide (H 2 O 2 ).
Light-emitting diodes (LEDs) demonstrate therapeutic effects for a range of biomedical applications including photodisinfection. Specific wavelength bands (centred at 405nm) are reported to be the most antimicrobial, however there remains no consensus on most effective irradiation parameters for optimal photodisinfection. The aim of this study was to assess decontamination efficiency by direct photodisinfection of monomicrobial biofilms using single (SWA), and multi wavelength (MWA) violet-blue light (VBL) arrays. Mature biofilms of nosocomial bacteria ( Acinetobacter baumannii , Pseudomonas aeruginosa , Escherichia coli, and Staphylococcus aureu s) were grown on 96 well polypropylene PCR plates. The biofilms were then exposed to VBL for 27.
Proliferation of microcystin (MC) producing Microcystis aeruginosa in brackish waters has been described in several locations and represents a new concern for public and environmental health. While the impact of a sudden salinity increase on M. aeruginosa physiology has been studied, less is known about the mechanisms involved in salt tolerance after acclimation. This study aims to compare the physiological responses of two strains of M. aeruginosa (PCC 7820 and PCC 7806), isolated from contrasted environments, to increasing salinities. After acclimation, growth, MC production and metabolomic analyses were conducted. For both strains, salinity decreased the biovolume, the growth and MC production rates and induced the accumulation of polyun.
Exposure to fungal spores has been associated with respiratory symptoms and allergic alveolitis among sawmill workers, but the complexity of sawmill workers' fungal exposure has been poorly studied. We characterized the fungal diversity in air samples from sawmill workers' breathing zone, and identified differences in richness, diversity and taxonomic composition between companies, departments, wood types and seasons.
The frequency and intensity of cyanobacterial blooms are increasing worldwide. Interactions between toxic cyanobacteria and aquatic microorganisms need to be critically evaluated to understand microbial drivers and modulators of the blooms. In this study, we applied 16S/18S rRNA gene sequencing and metabolomics analyses to measure the microbial community composition and metabolic responses of the cyanobacterium Microcystis aeruginosa in a coculture system receiving dissolved inorganic nitrogen and phosphorus (DIP) close to representative concentrations in Lake Taihu, China. M. aeruginosa secreted alkaline phosphatase using a DIP source produced by moribund and decaying microorganisms when the P source was insufficient. During this process,
The present study aimed to isolate Aeromonas from fish sold in the markets as well as in sushi and seafood shops and compare their virulence factors and antimicrobial characteristics with those of clinical isolates. Among the 128 fish isolates and 47 clinical isolates, A. caviae , A. dhakensis , and A. veronii were the principal species. A. dhakensis isolates carried at least 5 virulence genes, more than other Aeromonas species. The predominant genotype of virulence genes was hlyA/lip/alt/col/el in both A. dhakensis and A. hydrophila isolates, alt/col/ela in A. caviae isolates, and act in A. veronii isolates. A. dhakensis , A. hydrophila , and A. veronii isolates more often exhibited hemolytic and proteolytic activity and showed greater vir.
Bacterial biofilms can form persistent infections on wounds, on implanted medical devices, and are associated with many chronic diseases, such as cystic fibrosis. These infections are medically difficult to treat as biofilms are more resistant to antibiotic attack compared to their planktonic counterparts. The understanding of spatial and temporal variation in metabolism of biofilms is a critical component toward improved biofilm treatments. To this end, we developed oxygen-sensitive luminescent nanosensors to measure 3D oxygen gradients, an application of which is demonstrated here with Pseudomonas aeruginosa biofilms. The method was applied here and improves on traditional 1D methods of measuring oxygen profiles by investigating the spati.
Ca. Nitrosotenuis uzonensis is the only cultured moderately thermophilic member of the thaumarchaeotal order Nitrosopumilales (NP) that contains many mesophilic marine strains. We examined its membrane lipid composition at different growth temperatures (37, 46 and 50 °C). Its lipids were all membrane-spanning glycerol dialkyl glycerol tetraethers (GDGTs), with 0 to 4 cyclopentane moieties. Crenarchaeol (cren), the characteristic thaumarchaeotal GDGT, and its isomer (cren’) were present in high abundance (30-70 %). The GDGT polar headgroups were mono-, di- and trihexoses and hexose/phosphohexose. The ratio of glycolipid to phospholipid GDGTs was highest in the cultures grown at 50 °C. With increasing growth temperature, the relative contribu.
This study provides a phylogeographic insight into the population diversity of Xanthomonas translucens strains causing bacterial leaf streak disease of small-grain cereals in Iran. Among the 65 bacterial strains isolated from wheat, barley, and gramineous weeds in eight Iranian provinces, using multilocus sequence analyses and typing (MLSA/MLST) of four housekeeping genes ( i.e. dnaK , fyuA , gyrB , and rpoD ), 57 strains were identified as X. translucens pv. undulosa , while eight strains were identified as X. translucens pv. translucens . Although the pathogenicity patterns on oat and ryegrass weed species were varied among the strains, all X. translucens pv. undulosa strains were pathogenic on barley, Harding's grass, rye (except for XtK.
Lonely Guy (LOG) proteins are important enzymes in cellular organisms, which catalyze the final step in the production of biologically active cytokinins via dephosphoribosylation. LOG proteins are vital enzymes in plants for the activation of cytokinin precursors, which is crucial for plant growth and development. In fungi and bacteria, LOGs are implicated in pathogenic or non-pathogenic interactions with their plant hosts. However, LOGs have also been identified in the human pathogen Mycobacterium tuberculosis , and accumulation of cytokinin-degraded products, aldehydes, within bacterial cells is lethal to the bacterium in the presence of nitric oxide, suggesting diverse roles of LOGs in various species. In this study, we conducted biochem.
Liberibacter crescens is the only cultured member of its genus, which includes the devastating plant pathogen Ca. L. asiaticus ( C Las), associated with citrus greening/Huanglongbing (HLB). L. crescens has a larger genome and greater metabolic flexibility than C Las and the other uncultured plant pathogenic Liberibacter species, and it is currently the best model organism available for these pathogens. L. crescens grows slowly and dies rapidly under current culture protocols and this extreme fastidiousness makes it challenging to study. We have determined that a major cause of rapid death of L. crescens in batch culture is its alkalinization of the medium (to pH 8.5 by the end of logarithmic phase). The majority of this alkalinization is du.

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