Pseudomonas Putida Strain Research Articles

Mechanisms of Heavy Metal Tolerance in Bacteria: A Review

Heavy metal pollution from industrial activities and poor waste disposal poses significant environmental and health threats to humans and animals. This calls for sustainable approaches to the cleanup of heavy metals. This review explores metal tolerance mechanisms of bacteria such as the formation of biofilms, efflux systems, and enzymatic detoxification. These mechanisms allow bacteria communities to adapt and survive in contaminated environments. These adaptations are enhanced by mutations in the bacteria genes and by horizontal gene transfers, enabling bacteria species to survive under environmental stress while simultaneously contributing to nutrient cycling and the decomposition of organic matter. This review further explores the symbiotic interactions between bacteria, plants, and animals. These relationships enhance the metal tolerance ability of the different living organisms involved and are also very important in the bioremediation and phytoremediation of heavy metals. Plant growth-promoting rhizobacteria, Rhizobium, and Bacillus species are very important contributors to phytoremediation; they improve heavy metal uptake, improve the growth of roots, and plants resilience to stress. Moreover, this review highlights the importance of genetically engineered bacteria in closed-loop systems for optimized metal recovery. This offers environmentally friendly and sustainable options to the traditional remediation methods. Engineered Cupriavidus metallidurans CH34 and Pseudomonas putida strain 15420352 overexpressing metallothioneins have shown enhanced metal-binding capabilities, which makes them very effective in the treatment of industrial wastewaters and in biosorption applications. The use of engineered bacteria for the cleanup of heavy metals in closed-loop systems promotes the idea of a circular economy by recycling metals, thus reducing environmental waste. Multidisciplinary research that integrates synthetic biology, microbial ecology, and environmental science is very important for the advancement of metal bioremediation technologies. This review’s analysis on bacterial metal tolerance, symbiosis, and bioengineering strategies offers a pathway to effective bioremediation options, for the reclamation of heavy metal-polluted environments while promoting sustainable environmental practices.

Two-step biocatalytic conversion of post-consumer polyethylene terephthalate into value-added products facilitated by genetic and bioprocess engineering

Solving the plastic crisis requires high recycling quotas and technologies that allow open loop recycling. Here a biological plastic valorization approach consisting of tandem enzymatic hydrolysis and monomer conversion of post-consumer polyethylene terephthalate into value-added products is presented. Hydrolysates obtained from enzymatic degradation of pre-treated post-consumer polyethylene terephthalate bottles in a stirred-tank reactor served as the carbon source for a batch fermentation with an engineered Pseudomonas putida strain to produce 90mg/L of the biopolymer cyanophycin. Through fed-batch operation, the fermentation could be intensified to 1.4 g/L cyanophycin. Additionally, the upcycling of polyethylene terephthalate monomers to the biosurfactants (hydroxyalkanoyloxy)alkanoates and rhamnolipids is presented. These biodegradable products hold significant potential for applications in areas such as detergents, building blocks for novel polymers, and tissue engineering. In summary, the presented bio-valorization process underscores that addressing challenges like the plastic crisis requires an interdisciplinary approach.

Enhanced biosynthesis of poly(3-hydroxybutyrate) in engineered strains of Pseudomonas putida via increased malonyl-CoA availability.

Malonyl-coenzyme A (CoA) is a key precursor for the biosynthesis of multiple value-added compounds by microbial cell factories, including polyketides, carboxylic acids, biofuels, and polyhydroxyalkanoates. Owing to its role as a metabolic hub, malonyl-CoA availability is limited by competition in several essential metabolic pathways. To address this limitation, we modified a genome-reduced Pseudomonas putida strain to increase acetyl-CoA carboxylation while limiting malonyl-CoA utilization. Genes involved in sugar catabolism and its regulation, the tricarboxylic acid (TCA) cycle, and fatty acid biosynthesis were knocked-out in specific combinations towards increasing the malonyl-CoA pool. An enzyme-coupled biosensor, based on the rppA gene, was employed to monitor malonyl-CoA levels invivo. RppA is a type III polyketide synthase that converts malonyl-CoA into flaviolin, a red-colored polyketide. We isolated strains displaying enhanced malonyl-CoA availability via a colorimetric screening method based on the RppA-dependent red pigmentation; direct flaviolin quantification identified four engineered strains had a significant increase in malonyl-CoA levels. We further modified these strains by adding a non-canonical pathway that uses malonyl-CoA as precursor for poly(3-hydroxybutyrate) biosynthesis. These manipulations led to increased polymer accumulation in the fully engineered strains, validating our general strategy to boost the output of malonyl-CoA-dependent pathways in P. putida.

Draft genome sequencing of multidrug-resistant Pseudomonas putida strains, isolated from dairy cows with clinical mastitis and their farm environment.

We sequenced the genomes of three multidrug-resistant Pseudomonas putida strains namely 11CM-M1, 11CM-F1, and 11CM-S1, isolated from milk, feces, and farm soil samples collected from dairy cows with clinical mastitis. The assembled draft genomes of these P. putida strains were approximately 5.6 Mbp, with a GC content of 62.4%.

Production of bio-based adipic acid using a combination of engineered Pseudomonas putida strains

Adipic acid is an industrially important dicarboxylic acid and an essential precursor in the synthesis of Nylon-6,6. Due to the environmental impact associated with the current synthetic methods for Nylon-6,6 production, greener alternatives based on renewable and bio-based feedstocks are needed. In the present study, the potential of two Pseudomonas putida strains engineered for muconic acid accumulation was used to produce adipic acid from the lignin-derived aromatic compound guaiacol. This was achieved by expressing genes enabling the conversion of guaiacol to catechol in the first strain, and the conversion of muconic acid to adipic acid via expression of the Bacillus coagulans enoate reductase gene in the second strain. Through process engineering, a 2-step production was designed in which 2.93 mM bio-adipic acid was produced, with a yield of 0.57 mol/mol. This is the first demonstration that process and strain engineering can be combined to obtain a complete bio-based route for adipic acid production from depolymerized lignin compounds using the robust P. putida chassis.

Feed supplementation with quorum quenching probiotics improved growth response, immune response, and resistance in the giant mottled eel, Anguilla marmorata

This study aimed to evaluate the effect of feed supplementation with quorum quenching (QQ) probiotic bacteria on growth performance, hematology, immune response, and disease resistance in giant mottled eels. The feed used was supplemented with 45 ± 2.3 g/individual QQ bacteria using the following treatments, namely Bacillus cereus strain B8, Pseudomonas putida strain BPA1, Bacillus cereus strain HPC 576, and control (no QQ). A total of 420 eels (average weight 45 ± 2.3 g/ind.) were randomly reared in 12 open tarpaulin ponds of 2 x 1 x 1 (W x L x D) m and 1000-L volume for 8 weeks. Feed was given three times per day at 5% of the body weight. Water was changed daily at 25%, and shelters were provided using 3-inch diameter PVC pipes. The results showed that the feed containing QQ bacteria improved the growth, hematology, immunity, and disease resistance of the eels compared to the control (P < 0.05). Furthermore, among the QQ bacteria tested, the feed with B. cereus strain HPC 576 had the highest performance in all tested parameters.

Mitigating the detrimental impacts of low- and high-density polyethylene microplastics using a novel microbial consortium on a soil-plant system: Insights and interactions

The accumulation of polyethylene microplastics (PE-MPs) in soil has raised considerable concerns; however, the effects of their persistence and mitigation on agroecosystems have not been explored. This study aimed to assess the detrimental effects of PE-MPs on a soil-plant system and evaluate their mitigation using a novel microbial consortium (MC). We incorporated low-density polyethylene (LDPE) and high-density polyethylene (HDPE) at two different concentrations, along with a control (0 %, 1 %, and 2 % w/w) into the sandy loam soil for a duration of 135 days. The samples were also treated with a novel MC and incubated for 135 days. The MC comprised three bacterial strains (Ralstonia pickettii (MW290933) strain SHAn2, Pseudomonas putida strain ShA, and Lysinibacillus xylanilyticus XDB9 (T) strain S7–10F), and a fungal strain (Aspergillus niger strain F1–16S). Sunflowers were subsequently cultivated, and physiological growth parameters were measured. The results showed that adding 2 % LDPE significantly decreased soil pH by 1.06 units compared to the control. Moreover, adding 2 % HDPE resulted in a more significant decrease in soil electrical conductivity (EC) relative to LDPE and the control. A dose-dependent increase in dissolved organic carbon (DOC) was observed, with the highest DOC found in 2 % LDPE. The addition of higher dosages of LDPE reduced soil bulk density (BD) more than HDPE. The addition of 2 % HDPE increased the water drop penetration time (WDPT) but decreased the mean weight diameter of soil aggregates (MWD) and water-stable aggregates (WSA) compared to LDPE. The results also revealed that higher levels of LDPE enhanced soil basal respiration (BR) and microbial carbon biomass (MBC). The interaction of MC and higher MP percentages considerably reduced soil pH, EC, BD, and WDPT but significantly increased soil DOC, MWD, WSA, BR, and MBC. Regarding plant growth, incorporating 2 % PE-MPs significantly reduced physiological responses of sunflower: chlorophyll content (Chl; −15.2 %), Fv/Fm ratio (-25.3 %), shoot dry weight (ShD; −31.3 %), root dry weight (RD; −40 %), leaf area (LA; −38.4 %), and stem diameter (StemD; −25 %) compared to the control; however, the addition of novel MC considerably reduced and ameliorated the harmful effects of 2 % PE-MPs on the investigated plant growth responses.

Plastic-degrading microbial communities reveal novel microorganisms, pathways, and biocatalysts for polymer degradation and bioplastic production

Plastics derived from fossil fuels are used ubiquitously owing to their exceptional physicochemical characteristics. However, the extensive and short-term use of plastics has caused environmental challenges. The biotechnological plastic conversion can help address the challenges related to plastic pollution, offering sustainable alternatives that can operate using bioeconomic concepts and promote socioeconomic benefits. In this context, using soil from a plastic-contaminated landfill, two consortia were established (ConsPlastic-A and -B) displaying versatility in developing and consuming polyethylene or polyethylene terephthalate as the carbon source of nutrition. The ConsPlastic-A and -B metagenomic sequencing, taxonomic profiling, and the reconstruction of 79 draft bacterial genomes significantly expanded the knowledge of plastic-degrading microorganisms and enzymes, disclosing novel taxonomic groups associated with polymer degradation. The microbial consortium was utilized to obtain a novel Pseudomonas putida strain (BR4), presenting a striking metabolic arsenal for aromatic compound degradation and assimilation, confirmed by genomic analyses. The BR4 displays the inherent capacity to degrade polyethylene terephthalate (PET) and produce polyhydroxybutyrate (PHB) containing hydroxyvalerate (HV) units that contribute to enhanced copolymer properties, such as increased flexibility and resistance to breakage, compared with pure PHB. Therefore, BR4 is a promising strain for developing a bioconsolidated plastic depolymerization and upcycling process. Collectively, our study provides insights that may extend beyond the artificial ecosystems established during our experiments and supports future strategies for effectively decomposing and valorizing plastic waste. Furthermore, the functional genomic analysis described herein serves as a valuable guide for elucidating the genetic potential of microbial communities and microorganisms in plastic deconstruction and upcycling.

Adsorption of Flavonoids in a Transcriptional Regulator TtgR: Relative Binding Free Energies and Intermolecular Interactions.

Antimicrobial resistance in bacteria often arises from their ability to actively identify and expel toxic compounds. The bacterium strain Pseudomonas putida DOT-T1E utilizes its TtgABC efflux pump to confer robust resistance against antibiotics, flavonoids, and organic solvents. This resistance mechanism is intricately regulated at the transcriptional level by the TtgR protein. Through molecular dynamics and alchemical free energy simulations, we systematically examine the binding of seven flavonoids and their derivatives with the TtgR transcriptional regulator. Our simulations reveal distinct binding geometries and free energies for the flavonoids in the active site of the protein, which are driven by a range of noncovalent forces encompassing van der Waals, electrostatic, and hydrogen bonding interactions. The interplay of molecular structures, substituent patterns, and intermolecular interactions effectively stabilizes the bound flavonoids, confining their movements within the TtgR binding pocket. These findings yield valuable insights into the molecular determinants that govern ligand recognition in TtgR and shed light on the mechanism of antimicrobial resistance in P. putida DOT-T1E.

Nitrogen removal characteristics and Cr(VI) tolerance mechanisms of heterotrophic nitrifying bacterium Pseudomonas putida strain LX1

To solve the severe inhibition effect of nitrification process by heavy metals, necessitating the discovery of heavy metals-resistant nitrifying bacteria. In this study, a highly efficient bacterium, Pseudomonas putida strain LX1, capable of heterotrophic nitrification–aerobic denitrification (HN-AD) and resistant to hexavalent chromium (Cr(VI)) was isolated from activated sludge. Strain LX1 exhibited excellent growth activity and achieved efficient NH4+-N removal rates (94.07%–83.36 %) under 0–25 mg/L Cr(VI) stress. The heterotrophic nitrification ability of strain LX1 was superior to that of aerobic denitrification, and there was no accumulation of nitrite or nitrate during HN-AD process. Moreover, Three-dimensional excitation-emission matrix and Fourier-transform infrared spectroscopy analysis indicated that strain LX1 secreted more extracellular polymeric substances with negatively charged functional groups to bind Cr(VI) and alleviate its toxicity. Enzymatic analysis showed that strain LX1 countered the oxidative damage of reactive oxygen species by promoting the activities of oxidative dismutase and catalase. Furthermore, transcriptomic analyses revealed that the upregulation of genes associated with heavy metal substrate transporter proteins and antioxidant systems in strain LX1 contributed to the resistance against the high biotoxicity caused by Cr(VI). This study is important for understanding the heavy metal response mechanisms of HN-AD strains and its potential application in practical wastewater treatment.

Potential biocontrol agents involved in induction of defense related enzymes in brinjal (solanum melongena l.) Against phomopsis blight caused by phomopsis vexans

Phomopsis blight, caused by Phomopsis vexans (P. vexans) in brinjal plants is one of the main diseases in India. In order to manage this malady, we initially screened seed samples of 14 cultivars obtained from seed suppliers for the disease incidence. Based on the disease incidence and severity, the samples were categorized as resistant and susceptible cultivars of brinjal. It was established that MEBH-9' was a sensitive cultivar, while the Kolar local was resistant to the disease. Two ideal strains of Rhizosphere colonizing bacteria (RCB), Pseudomonas putida strain Has-1/c (HM229805), and Phylloplane Colonizing Bacteria (PCB), Bacillus subtilis strain Br/ph-33 (KJ867501), previously characterized as potential plant growth-promoting rhizobacteria, were tested for their potential biocontrol of this disease. Both susceptible and resistant cultivars' seedlings were treated with different combinations of PCB and RCB, PCB alone, RCB alone, and challenge inoculated with P. vexans. In order to ascertain whether there was signalling between defense enzyme activation and plant protection after PCB and RCB treatment, defense enzymes were quantified. Brinjal plants treated with PCB+RCB combination and challenge inoculated with P. vexans showed a significant increase in the activity of Phenylalanine ammonia-lyase (PAL), Peroxidase (POX), Lipoxygenase (LOX), Polyphenol oxidase (PPO), Catalase, Chitinase, β -1,3-glucanase, and Hydrogen peroxide (H2O2) content. The current study reveals that the defense enzymes and PR-proteins are gradually induced and accumulated, which enhance the resistance in brinjal plants against P. vexans-causing fruit rot disease.

Analysis of microbial community evolution, autolysis phenomena, and energy metabolism pathways in Pholiota nameko endophytes.

Pholiota nameko is a widely consumed edible fungus. This study focuses on two crucial developmental stages of Pholiota nameko, namely, mycelium and ascospores. The objectives of this research were to investigate changes in microbial diversity and community structure during the growth of Pholiota nameko and to analyze the adaptability of the dominant strains to their respective habitats through metabolic. Specifically, we conducted second-generation sequencing of the 16S rRNA gene (Illumina) on samples obtained from these stages. In addition, we isolated and characterized endophytes present in Pholiota nameko, focusing on examining the impact of dominant endophyte genera on autolysis. We also conducted a metabolic pathway analysis. The results unveiled 578,414 valid sequences of Pholiota nameko endophytic fungi. At the phylum level, the dominant taxa were Basidiomycota, Ascomycota, Zoopagomycota, and Mucoromycota. At the genus level, the dominant taxa observed were Pholiota, Inocybe, Fusarium, and Hortiboletus. For endophytic bacteria, we obtained 458,475 valid sequences. The dominant phyla were Proteobacteria, TM6, Firmicutes, and Bacteroidetes, while the dominant genera were Edaphobacter, Xanthomonas, Burkholderia, and Pseudomonas. Moreover, we identified the isolated strains in Pholiota nameko using 16S rDNA, and most of them were found to belong to the genus Pseudomonas, with Pseudomonas putida being the most prevalent strain. The findings revealed that the Pseudomonas putida strain has the ability to slow down the breakdown of soluble proteins and partially suppress the metabolic processes that generate superoxide anion radicals in Pholiota nameko, thereby reducing autolysis. Additionally, our results demonstrated that molybdenum enzyme-mediated anaerobic oxidative phosphorylation reactions were the primary energy metabolism pathway in the Pseudomonas putida strain. This suggests that the molybdenum cofactor synthesis pathway might be the main mechanism through which Pholiota nameko adapts to its complex and diverse habitats.

Enhancing Botrytis disease management in tomato plants: insights from a Pseudomonas putida strain with biocontrol activity.

This study explores the biocontrol potential of Pseudomonas putida Z13 against Botrytis cinerea in tomato plants, addressing challenges posed by the pathogen's fungicide resistance. The aims of the study were to investigate the in vitro and in silico biocontrol traits of Z13, identify its plant-colonizing efficacy, evaluate the efficacy of different application strategies against B. cinerea in planta, and assess the capacity of Z13 to trigger induced systemic resistance (ISR) in plants. The in vitro experiments revealed that Z13 inhibits the growth of B. cinerea, produces siderophores, and exhibits swimming and swarming activity. Additionally, the Z13 genome harbors genes that encode compounds triggering ISR, such as pyoverdine and pyrroloquinoline quinone. The in planta experiments demonstrated Z13's efficacy in effectively colonizing the rhizosphere and leaves of tomato plants. Therefore, three application strategies of Z13 were evaluated against B. cinerea: root drenching, foliar spray, and the combination of root drenching and foliar spray. It was demonstrated that the most effective treatment of Z13 against B. cinerea was the combination of root drenching and foliar spray. Transcriptomic analysis showed that Z13 upregulates the expression of the plant defense-related genes PR1 and PIN2 upon B. cinerea inoculation. The results of the study demonstrated that Z13 possesses significant biocontrol traits, such as the production of siderophores, resulting in significant plant protection against B. cinerea when applied as a single treatment to the rhizosphere or in combination with leaf spraying. Additionally, it was shown that Z13 root colonization primes plant defenses against the pathogen.

QsdS lactonase from Sphingopyxis sp. strain EG6 inhibits biofilm formation by Pseudomonas putida strain TS312 by degrading N-acyl homoserine lactone

Inhibiting biofilm formation triggered by a quorum sensing (QS) mechanism is promising in industries where biofilms cause deterioration of production performance. QS can be disrupted by degrading or scavenging the signal compounds that mediate QS, such as N-acyl-homoserine lactones (AHLs). This study investigated the effect of lactonase, an AHL-degrading enzyme, on bacterial biofilm formation; we applied lactonase QsdS, produced by Sphingopyxis sp. strain EG6, which was isolated from a cooling water system, to biofilms of Pseudomonas putida strain TS312, which was isolated from white water in a paper mill. Addition of QsdS lactonase inhibited biofilm formation by P. putida strain TS312; the magnitude of the effect was Qsds lactonase dose-dependent, decreasing the biofilm mass by 85% at a Qsds lactonase concentration of 10 µg/mL. Addition of QsdS lactonase decreased AHL concentrations in the culture supernatant of P. putida strain TS312, indicating that degradation of AHLs inhibited biofilm formation by disrupting QS. Observing three-dimensional biofilms by confocal laser scanning microscopy (CLSM) and optical coherence tomography (OCT) showed that addition of QsdS lactonase decreased the secretion of α-polysaccharides by P. putida strain TS312.

Comparison of wild-type KT2440 and genome-reduced EM42 Pseudomonas putida strains for muconate production from aromatic compounds and glucose

Pseudomonas putida KT2440 is a robust, aromatic catabolic bacterium that has been widely engineered to convert bio-based and waste-based feedstocks to target products. Towards industrial domestication of P. putida KT2440, rational genome reduction has been previously conducted, resulting in P. putida strain EM42, which exhibited characteristics that could be advantageous for production strains. Here, we compared P. putida KT2440- and EM42-derived strains for cis,cis-muconic acid production from an aromatic compound, p-coumarate, and in separate strains, from glucose. To our surprise, the EM42-derived strains did not outperform the KT2440-derived strains in muconate production from either substrate. In bioreactor cultivations, KT2440- and EM42-derived strains produced muconate from p-coumarate at titers of 45 g/L and 37 g/L, respectively, and from glucose at 20 g/L and 13 g/L, respectively. To provide additional insights about the differences in the parent strains, we analyzed growth profiles of KT2440 and EM42 on aromatic compounds as the sole carbon and energy sources. In general, the EM42 strain exhibited reduced growth rates but shorter growth lags than KT2440. We also observed that EM42-derived strains resulted in higher growth rates on glucose compared to KT2440-derived strains, but only at the lowest glucose concentrations tested. Transcriptomics revealed that genome reduction in EM42 had global effects on transcript levels and showed that the EM42-derived strains that produce muconate from glucose exhibit reduced modulation of gene expression in response to changes in glucose concentrations. Overall, our results highlight that additional studies are warranted to understand the effects of genome reduction on microbial metabolism and physiology, especially when intended for use in production strains.

Autoxidation Catalysis for Carbon-Carbon Bond Cleavage in Lignin.

Selective lignin depolymerization is a key step in lignin valorization to value-added products, and there are multiple catalytic methods to cleave labile aryl-ether bonds in lignin. However, the overall aromatic monomer yield is inherently limited by refractory carbon-carbon linkages, which are abundant in lignin and remain intact during most selective lignin deconstruction processes. In this work, we demonstrate that a Co/Mn/Br-based catalytic autoxidation method promotes carbon-carbon bond cleavage in acetylated lignin oligomers produced from reductive catalytic fractionation. The oxidation products include acetyl vanillic acid and acetyl vanillin, which are ideal substrates for bioconversion. Using an engineered strain of Pseudomonas putida, we demonstrate the conversion of these aromatic monomers to cis,cis-muconic acid. Overall, this study demonstrates that autoxidation enables higher yields of bioavailable aromatic monomers, exceeding the limits set by ether-bond cleavage alone.

Mitigation of biogenic methanethiol using bacteriophages in synthetic wastewater augmented with Pseudomonas putida

Wastewater malodour is the proverbial ‘elephant in the room’ notwithstanding its severe implications on sanitation, health, and hygiene. The predominant malodorous compounds associated with wastewater treatment plants and toilets are volatile organic compounds, such as hydrogen sulphide, ammonia, methanethiol, and organic acids. Among them, methanethiol warrants more attention owing to its relatively low olfactory threshold and associated cytotoxicity. This requires an efficient odour-abatement method since conventional techniques are either cost-prohibitive or leave recalcitrant byproducts. Bacteriophage-based methodology holds promise, and the described work explores the potential. In this study, a non-lysogenous Pseudomonas putida strain is used as a model organism that produces methanethiol in the presence of methionine. Two double-stranded DNA phages of genome sizes > 10 Kb were isolated from sewage. ɸPh_PP01 and ɸPh_PP02 were stable at suboptimal pH, temperature, and at 10% chloroform. Moreover, they showed adsorption efficiencies of 53% and 89% in 12 min and burst sizes of 507 ± 187 and 105 ± 7 virions per cell, respectively. In augmented synthetic wastewater, ɸPh_PP01 and ɸPh_PP02 reduced methanethiol production by 52% and 47%, respectively, with the concomitant reduction in P. putida by 3 logs in 6 h. On extension of the study in P. putida spiked-sewage sample, maximum reduction in methanethiol production was achieved in 3 h, with 49% and 48% for ɸPh_PP01 and ɸPh_PP02, respectively. But at 6 h, efficiency reduced to 36% with both the phages. The study clearly demonstrates the potential of phages as biocontrol agents in the reduction of malodour in wastewater.

Enhancing earthworm (Lumbricus terrestris) tolerance to plastic contamination through gut microbiome fortification with plastic-degrading microorganisms

Microorganisms from L. terrestris gut previously exposed to different types of plastic (PET, LDPE, LLDPE, and PS) were studied to be used as probiotics of earthworms in plastic-contaminated soils (LDPE, LLDPE and recycled mulching film) at mesocosm-scale trials. The most abundant morphotypes with enzymatic capacities of interest were identified. Pseudomonas alkylphenolica (PL4) and Pseudomonas putida (PL5) strains were selected to be used as inoculants using Morus alba leaves as carriers to strengthen the intestinal microbiota of earthworms. Culture (selective cetrimide agar medium) and molecular (qPCR) techniques were used to trace the presence of the inoculum in the intestine of the earthworms. Additionally, a metataxonomic analysis was carried out to study the biodiversity and functionality of the earthworm microbiome, and their measure of survival and weight. Probiotics improved the survival rates of earthworms exposed to plastics, which also increased the abundance of microbial groups of interest in plastic bioremediation tasks.

Analysis of Pathogens of Urinary Tract Infections Associated with Indwelling Double-J Stents and Their Susceptibility to Globularia alypum.

Ureteral double-J stents are frequently used to prevent urinary obstruction. They can develop bacterial colonization and encrustation, which leads to persistent infections that seldom respond to antibiotic treatment. Thus, the goal of this study was to evaluate the local spectrum of bacterial pathogens and their susceptibility to natural compounds. A total of 59 double-J ureteral stents from 59 consecutive patients were examined. The samples were inoculated on agar culture mediums. Extracts of Globularia alypum L. were evaluated for their antibacterial activity with the diffusion and broth dilution methods; for antibiofilm activity, the crystal violet assay was used. The identification and the quantification of the different constituents of extracts were determined by reverse-phase high-performance liquid chromatography (RP-HPLC). Bacterial growth was found in three patients (5.1%). Enterococcus faecalis (1.7%), Acinetobacter baumanii (1.7%), and Pseudomonas putida (1.7%) strains were more commonly detected. They were resistant to several common antibiotics. All extracts presented several components, mainly nepetin-7-glucoside and trans-ferulic-acid, and they had antibacterial activity (MIC = 6.25 mg/mL and MBC = 6.25 mg/mL), and antibiofilm (59.70% at 25 mg/mL) properties, especially against Acinetobacter baumanii. The results achieved confirm the important role of this plant as a source of therapeutic activities.

Metabolome and transcriptome reprogramming underlying tomato drought resistance triggered by a Pseudomonas strain

Although amelioration of drought stress by Plant Growth Promoting Rhizobacteria (PGPR) is a well-documented phenomenon, the combined molecular and metabolic mechanisms governing this process remain unclear. In these lines, the present study aimed to provide new insights in the underlying drought attenuating mechanisms of tomato plants inoculated with a PGP Pseudomonas putida strain, by using a combination of metabolomic and transcriptomic approaches. Following Differentially Expressed Gene analysis, it became evident that inoculation resulted in a less disturbed plant transcriptome upon drought stress. Untargeted metabolomics highlighted the differential metabolite accumulation upon inoculation, as well as the less metabolic reprograming and the lower accumulation of stress-related metabolites for inoculated stressed plants. These findings were in line with morpho-physiological evidence of drought stress mitigation in the inoculated plants. The redox state modulation, the more efficient nitrogen assimilation, as well as the differential changes in amino acid metabolism, and the induction of the phenylpropanoid biosynthesis pathway, were the main drought-attenuating mechanisms in the SAESo11-inoculated plants. Shifts in pathways related to hormonal signaling were also evident upon inoculation at a transcript level and in conjunction with carbon metabolism regulation, possibly contributed to a drought-attenuation preconditioning. The identified signatory molecules of SAESo11-mediated priming against drought included aspartate, myo-inositol, glutamate, along with key genes related to trehalose, tryptophan and cysteine synthesis. Taken together, SAESo11-inoculation provides systemic effects encompassing both metabolic and regulatory functions, supporting both seedling growth and drought stress amelioration.