Pseudomonas Putida Research Articles

Exploring the Antimicrobial Properties of 99 Natural Flavour and Fragrance Raw Materials against Pathogenic Bacteria: A Comparative Study with Antibiotics

Currently, one of the most serious global problems is the increasing incidence of infectious diseases. This is closely related to the increase in antibiotic use, which has resulted in the development of multidrug resistance in microorganisms. Another problem is the numerous microbiological contaminations of cosmetic products, which can lead to dangerous bacterial infections in humans. Natural fragrance raw materials exhibit a wide spectrum of biological properties, including antimicrobial properties. Despite their prevalence and availability on the commercial market, there is little research into their effects on multidrug-resistant microorganisms. This study examines the inhibitory effect of natural substances on Gram-positive and Gram-negative bacteria. For this purpose, screening and appropriate assays were carried out to determine the minimum inhibitory concentration (MIC) value of individual substances, using the alamarBlueTM reagent. The lowest MIC values were observed for Staphylococcus aureus (black seed (Nigella sativa) expressed oil, MIC = 25 µg/mL), Kocuria rhizophila (fir balsam absolute, MIC = 12.5 µg/mL), and Pseudomonas putida (cubeb oil and fir balsam absolute, MIC = 12.5 µg/mL). The most resistant Gram-negative species was Enterobacter gergoviae, while Staphylococcus epidermidis was the most resistant Gram-positive species.

The phosphodiesterase DibA interacts with the c-di-GMP receptor LapD and specifically regulates biofilm in Pseudomonas putida.

The ubiquitous bacterial second messenger c-di-GMP is synthesized by diguanylate cyclase and degraded by c-di-GMP-specific phosphodiesterase. The genome of Pseudomonas putida contains dozens of genes encoding diguanylate cyclase/phosphodiesterase, but the phenotypical-genotypical correlation and functional mechanism of these genes are largely unknown. Herein, we characterize the function and mechanism of a P. putida phosphodiesterase named DibA. DibA consists of a PAS domain, a GGDEF domain, and an EAL domain. The EAL domain is active and confers DibA phosphodiesterase activity. The GGDEF domain is inactive, but it promotes the phosphodiesterase activity of the EAL domain via binding GTP. Regarding phenotypic regulation, DibA modulates the cell surface adhesin LapA level in a c-di-GMP receptor LapD-dependent manner, thereby inhibiting biofilm formation. Moreover, DibA interacts and colocalizes with LapD in the cell membrane, and the interaction between DibA and LapD promotes the PDE activity of DibA. Besides, except for interacting with DibA and LapD itself, LapD is found to interact with 11 different potential diguanylate cyclases/phosphodiesterases in P. putida, including the conserved phosphodiesterase BifA. Overall, our findings demonstrate the functional mechanism by which DibA regulates biofilm formation and expand the understanding of the LapD-mediated c-di-GMP signaling network in P. putida.

Metagenomics harvested genus-specific single-stranded DNA-annealing proteins improve and expand recombineering in Pseudomonas species.

The widespread Pseudomonas genus comprises a collection of related species with remarkable abilities to degrade plastics and polluted wastes and to produce a broad setof valuable compounds, ranging from bulk chemicals to pharmaceuticals. Pseudomonas possess characteristics of tolerance and stress resistance making them valuable hosts for industrial and environmental biotechnology. However, efficient and high-throughput genetic engineering tools have limited metabolic engineering efforts and applications. To improve their genome editing capabilities, we firstemployed a computational biology workflow to generate a genus-specific library of potential single-stranded DNA-annealing proteins (SSAPs). Assessment of the library was performed in different Pseudomonas using a high-throughput pooled recombinase screen followed by Oxford Nanopore NGS analysis. Among different active variants with variable levels of allelic replacement frequency (ARF), efficient SSAPs were found and characterized for mediating recombineering in the four tested species. New variants yielded higher ARFs than existing ones in Pseudomonas putida andPseudomonas aeruginosa, andexpanded the field of recombineering in Pseudomonas taiwanensisand Pseudomonas fluorescens. These findings will enhance the mutagenesis capabilities of these members of the Pseudomonas genus, increasing the possibilities for biotransformation and enhancingtheir potential for synthetic biology applications. .

Transition of stability of putidaredoxin reductase by introducing proline

The monooxygenase activity of the Pseudomonas putida cytochrome P450 system was expressed in the presence of P450cam, putidaredoxin reductase (PdR), and putidaredoxin (Pd). In turn, it has been found to catalyze the oxidation of various organic compounds. Since cytochrome P450 system substrates are often insoluble in water, reactions should be carried out at high temperatures and in the presence of organic solvents where the high stability of all of P450cam, PdR, and Pd are needed. In this study, proline was introduced to improve the stability of PdR which was estimated to have the lowest organic solvent stability compared to others. Thus, the half-life of PdR_T221P at 40°C was 1.37 times longer than that of wild-type PdR. However, the half-life of PdR_T221P in the presence of 25% (v/v) methanol was shorter than that of wild-type PdR. Furthermore, structural stabilities of PdRs using molecular dynamics (MD) simulations demonstrated these phenomena.

Metabolic impact of heterologous protein production in Pseudomonas putida: Insights into carbon and energy flux control

For engineered microorganisms, the production of heterologous proteins that are often useless to host cells represents a burden on resources, which have to be shared with normal cellular processes. Within a certain metabolic leeway, this competitive process has no impact on growth. However, once this leeway, or free capacity, is fully utilized, the extra load becomes a metabolic burden that inhibits cellular processes and triggers a broad cellular response, reducing cell growth and often hindering the production of heterologous proteins. In this study, we sought to characterize the metabolic rearrangements occurring in the central metabolism of Pseudomonas putida at different levels of metabolic load. To this end, we constructed a P. putida KT2440 strain that expressed two genes encoding fluorescent proteins, one in the genome under constitutive expression to monitor the free capacity, and the other on an inducible plasmid to probe heterologous protein production. We found that metabolic fluxes are considerably reshuffled, especially at the level of periplasmic pathways, as soon as the metabolic load exceeds the free capacity. Heterologous protein production leads to the decoupling of anabolism and catabolism, resulting in large excess energy production relative to the requirements of protein biosynthesis. Finally, heterologous protein production was found to exert a stronger control on carbon fluxes than on energy fluxes, indicating that the flexible nature of P. putida's central metabolic network is solicited to sustain energy production.

Improved biodegradability of low-density polyethylene using plasma pretreatment and plastic-degrading bacteria

Polyethylene is widely used and is a major cause of serious environmental problems because it is non-biodegradable and difficult to recycle. To overcome these environmental problems, in this study, low-density polyethylene (LDPE) was treated with low-temperature plasma at atmospheric pressure, an eco-friendly technology. After inducing oxidation on the LDPE film by plasma treatment, a method for improving the biodegradation efficiency of LDPE-degrading bacteria was identified, and the underlying mechanism was investigated. Fourier-transform infrared and X-ray photoelectron spectroscopy analyses showed that the plasma treatment resulted in the formation of new peaks corresponding to the C–O and –CC– bonds and stronger –CO– bond groups than the –C–O– bond groups, showing that the oxidation of LDPE film was strong. Additionally, the –C–H– bond ratio on LDPE surfaces inoculated with Pseudomonas putida increased by 13–25.5% compared to that on untreated controls. Pseudomonas putida reduced the thermal stability by forming a biofilm on the oxidized LDPE film during the plasma treatment. The weight of untreated LDPE film was reduced by 50%, while that of treated LDPE film was reduced by 80%, confirming more efficient degradation. Thus, LDPE decomposition efficiency can be improved using a short plasma treatment to increase the adhesion and fixation of LDPE-degrading bacteria on the surface of oxidized LDPE films. Therefore, plasma pretreatment technology could be used to reduce plastic contamination in agricultural environments.

Heterologous constitutive production of short-chain-length polyhydroxyalkanoates in Pseudomonas putida KT2440: the involvement of IbpA inclusion body protein.

Designing cell factories for the production of novel polyhydroxyalkanoates (PHAs) via smart metabolic engineering is key to obtain à la carte materials with tailored physicochemical properties. To this end, we used the model medium-chain-length-PHA producing bacterium, P. putida KT2440 as a chassis, which is characterized by its metabolic versatility and stress tolerance. Different PHA biosynthetic modules were assembled in expression plasmids using the Golden gate/MoClo modular assembly technique to implement an orthogonal short-chain-lengh-PHA (scl-PHA) switch in a "deaf" PHA mutant. This was specifically constructed to override endogenous multilevel regulation of PHA synthesis in the native strain. We generated a panel of engineered approaches carrying the genes from Rhodospirillum rubrum, Cupriavidus necator and Pseudomonas pseudoalcaligenes, demonstrating that diverse scl-PHAs can be constitutively produced in the chassis strain to varying yields from 23% to 84% PHA/CDW. Co-feeding assays of the most promising engineered strain harboring the PHA machinery from C. necator resulted to a panel of PHBV from 0.6% to 19% C5 monomeric incorporation. Chromosomally integrated PHA machineries with high PhaCCn synthase dosage successfully resulted in 68% PHA/CDW production. Interestingly, an inverse relationship between PhaC synthase dosage and granule size distribution was demonstrated in the heterologous host. In this vein, it is proposed the key involvement of inclusion body protein IbpA to the heterologous production of tailored PHA in P. putida KT2440.

Mathematical modeling for operative improvement of the decoloration of Acid Red 27 by a novel microbial consortium of Trametes versicolor and Pseudomonas putida: A multivariate sensitivity analysis

In this work, it is presented a first approach of a mathematical and kinetic analysis for improving the decoloration and further degradation process of an azo dye named acid red 27 (AR27), by means of a novel microbial consortium formed by the fungus Trametes versicolor and the bacterium Pseudomonas putida. A multivariate analysis was carried out by simulating scenarios with different operating conditions and developing a specific mathematical model based on kinetic equations describing all stages of the biological process, from microbial growth and substrate consuming to decoloration and degradation of intermediate compounds. Additionally, a sensitivity analysis was performed by using a factorial design and the Response Surface Method (RSM), for determining individual and interactive effects of variables like, initial glucose concentration, initial dye concentration and the moment in time for bacterial inoculation, on response variables assessed in terms of the minimum time for: full decoloration of AR27 (R1 = 2.375 days); maximum production of aromatic metabolites (R2 = 1.575 days); and full depletion of aromatic metabolites (R3 = 12.9 days). Using RSM the following conditions improved the biological process, being: an initial glucose concentration of 20 g l-1, an initial AR27 concentration of 0.2 g l-1 and an inoculation moment in time of P. putida at day 1. The mathematical model is a feasible tool for describing AR27 decoloration and its further degradation by the microbial consortium of T. versicolor and P. putida, this model will also work as a mathematical basis for designing novel bio-reaction systems than can operate with the same principle of the described consortium.

The metagenomic and metabolomic profile of the gut microbes in Chinese full-term and late preterm infants treated with Clostridium butyricum

The present study investigated the composition, abundance, and diversity of gut microbes in full-term and late-preterm infants from a medical center in eastern China. A total of 144 genomes of stool samples were captured for 16S rRNA metagenomic analyses. A high abundance of commensal intestinal bacteria was detected in these samples such as Phocaeicola vulgatus, Escherichia coli, and Faecalibacterium prausnitzii, indicating a relatively consistent diversity of gut microbes in the present full-term infants aged 38–40 weeks. However, late preterm infants (n = 50) with mandatory antimicrobials feeding exhibited lower diversity but a higher composition of opportunistic pathogens such as Enterococcus species. Centralized on the situation, we explored the regulatory effect of Clostridium butyricum as probiotics on these late preterm infants. The consumption of C. butyricum did not restore the composition of gut microbes altered by antimicrobials to normal levels, although several opportunistic pathogens decreased significantly after probiotic therapy including Staphylococcus aureus, Sphingomonas echinoides, and Pseudomonas putida. We also compared the effects of day-fed versus night-fed probiotics. Intriguingly, the nighttime feeding showed a higher proportion of C. butyricum compared with probiotic day-feeding. Finally, fecal metabolome and metabolites were analyzed in late preterm infants with (n = 20) or without probiotic therapy (n = 20). The KEGG enrichment analysis demonstrated that vitamin digestion and absorption, synaptic vesicle cycle, and biotin metabolism were significantly increased in the probiotic-treated group, while MSEA indicated that a series of metabolism were significantly enriched in probiotic-treated infants including glycerolipid, biotin, and lysine, indicating the complex effects of probiotic therapy on glutathione metabolism and nutrients digestion and absorption in late preterm infants. Overall, this study provided metagenomic and metabolomic profile of the gut microbes in full-term newborns and late preterm infants in eastern China. Further studies are needed to support and elucidate the role of probiotic feeding in late preterm infants with mandatory antimicrobial treatment.

A transcription factor-based bacterial biosensor system and its application for on-site detection of explosives

Detecting unexploded landmines is critical due to the environmental pollution and potential humanitarian risks caused by buried landmines. Therefore, this study focused on developing a biosensor system capable of detecting explosives safely and efficiently. A novel transcription factor-based Escherichia coli biosensor was designed to detect 1,3-dinitrobenzene (1,3-DNB). The MexT transcription factor from Pseudomonas putida (P. putida) was identified as the fundamental sensing element in this biosensor. The study found that MexT positively regulated the transcription of PP_2827 by binding to the bidirectional promoter region between them, and significantly enhanced the expression of downstream genes under the condition of 1,3-DNB. The MexT-based biosensor for 1,3-DNB was developed by adopting different combinations of the mexT gene and promoters. The optimized biosensor demonstrated adequate sensitivity for detecting 0.1 μg/mL of 1,3-DNB in a liquid solution with satisfactory specificity and long-term stability. Subsequently, the MexT-based biosensor was integrated into a detection device to simulate the in-field exploration of explosives. The system exhibited a detection sensitivity of 0.5 mg/kg for 1,3-DNB in the sand, and realized the detection of on-site and large-scale area and the location of buried 1,3-DNB under the soil. The study provided a novel transcription factor-based bacterial biosensor and a complete system (China Earth Eye, CEE) for sensitive detection of 1,3-DNB. The good performance of this biosensor system can facilitate the development of accurate, on-site, and high-efficient exploration of explosives in real extensive minefields. Moreover, this 1,3-DNB biosensor can be complementary to the 2,4-DNT biosensor reported before, demonstrating its potential applications in military situations.

Isolation, screening, and identification of chitinase-producing bacterial strains from riverbank soils at Ambo, Western Ethiopia

Chitinases are hydrolytic enzymes that dissolve the glycosidic linkages in chitin. Chitin is a cell wall component of fungi and fund in exoskeleten of worms and arthropods. Chitinase has been applied in agriculture, as a biopesticide for the control of plant fungal infections, in medicine, and in waste management. This research aimed to isolate, screen, and identification of chitinase-producing bacteria from riverbank soils. Twenty nine chitinolytic bacteria were isolated from the river bank soil samples, from which 9 of them had strong chitinolytic properties. Chitinase production was determined by zones of hydrolysis produced after 96 h of incubation at 37 °C. The different bacterial isolates were characterized morphologically, microscopically, and biochemically and finally eight strain were identified at species level by Matrix Assisted Laser Desorption Ionization - Time of Flight Mass Spectrometry (MALDI–TOF MS). From the eight, bacterial isolates investigated in this study Stenotrophomonas maltophilia showed the highest chitinase enzyme activity (625 μg/mL) followed by Pseudomonas putida with the enzyme activity of (553 μg/mL) and the least enzyme activity was recorded for Lilliottia amnigena (80 μg/mL). An incubation temperature of 45 °C, neutral pH and an incubation period of 96 h are found to be the optimum condition for the chitinase enzyme production from Stenotrophomonas maltophilia. The results of this study indicated the possibility of the production of chitinase from the chitinolytic bacterial isolates, which was highly useful for a variety of applications, including biocontrol of harmful insects and pathogenic fungi as well as in the biochemical, pharmaceutical, and medical sectors.

The Inhibitory Effects of Amylase and Streptokinase on Minimum Inhibitory Concentration of Antibiotics Used to Treat Gram Negative Bacteria Biofilm Infection on Indwelling Devices.

The study evaluated and compared the effect of adding streptokinase and amylase to antibiotics that are already used in clinical practice to treat Gram negative bacteria biofilm infection on indwelling devices on the antibiotics' minimum inhibitory concentration (MIC). 24h-old biofilms were developed on 96-well plate with eight clinical isolates. MIC of amikacin, cefepime, ceftazidime, colistin, meropenem, and piperacillin-tazobactam, on biofilms were measured before and after the addition of 25U/ml streptokinase and 25μg/ml amylase with microplate reader. The addition of streptokinase reduces the MICs of cefepime, ceftazidime, colistin, meropenem from (16, 16, 8, 4μg/ml) to (8, 1, 1, 0.5μg/ml) in Escherichia coli (isolate 1). While the addition of amylase reduces the MICs of only cefepime, ceftazidime from (16, 16μg/ml) to (2, 4μg/ml) in E. coli (isolate 1). In Pseudomonas aeruginosa (isolate 4), the MICs of amikacin, cefepime, ceftazidime, colistin and meropenem (64, 16, 32, 4, 32μg/ml) reduced to (2, 1, 0.5, 0.25, 0.5μg/ml) with streptokinase and (4, 4, 4, 2, 0.5μg/ml) with amylase respectively. Similar inhibitions were seen in Pseudomonas putida, Proteus mirabilis. We can conclude that the addition of streptokinase and amylase were effective in reducing the MICs of antibiotics that are commonly used to treat Gram negative bacteria biofilm infection on indwelling devices, thereby increasing susceptibility of bacteria to antibiotics. Streptokinase obviously had a greater effect than amylase, implying that it should be prioritized in future in vivo and clinical studies to obtain successful therapy with antibiotics on biofilm infections.

TudS desulfidases recycle 4-thiouridine-5’-monophosphate at a catalytic [4Fe-4S] cluster

In all domains of life, transfer RNAs (tRNAs) contain post-transcriptionally sulfur-modified nucleosides such as 2- and 4-thiouridine. We have previously reported that a recombinant [4Fe-4S] cluster-containing bacterial desulfidase (TudS) from an uncultured bacterium catalyzes the desulfuration of 2- and 4-thiouracil via a [4Fe-5S] cluster intermediate. However, the in vivo function of TudS enzymes has remained unclear and direct evidence for substrate binding to the [4Fe-4S] cluster during catalysis was lacking. Here, we provide kinetic evidence that 4-thiouridine-5’-monophosphate rather than sulfurated tRNA, thiouracil, thiouridine or 4-thiouridine-5’-triphosphate is the preferred substrate of TudS. The occurrence of sulfur- and substrate-bound catalytic intermediates was uncovered from the observed switch of the S = 3/2 spin state of the catalytic [4Fe-4S] cluster to a S = 1/2 spin state upon substrate addition. We show that a putative gene product from Pseudomonas putida KT2440 acts as a TudS desulfidase in vivo and conclude that TudS-like enzymes are widespread desulfidases involved in recycling and detoxifying tRNA-derived 4-thiouridine monophosphate nucleosides for RNA synthesis.

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.

Maximizing Heterologous Expression of Engineered Type I Polyketide Synthases: Investigating Codon Optimization Strategies.

Type I polyketide synthases (T1PKSs) hold enormous potential as a rational production platform for the biosynthesis of specialty chemicals. However, despite great progress in this field, the heterologous expression of PKSs remains a major challenge. One of the first measures to improve heterologous gene expression can be codon optimization. Although controversial, choosing the wrong codon optimization strategy can have detrimental effects on the protein and product levels. In this study, we analyzed 11 different codon variants of an engineered T1PKS and investigated in a systematic approach their influence on heterologous expression in Corynebacterium glutamicum, Escherichia coli, and Pseudomonas putida. Our best performing codon variants exhibited a minimum 50-fold increase in PKS protein levels, which also enabled the production of an unnatural polyketide in each of these hosts. Furthermore, we developed a free online tool (https://basebuddy.lbl.gov) that offers transparent and highly customizable codon optimization with up-to-date codon usage tables. In this work, we not only highlight the significance of codon optimization but also establish the groundwork for the high-throughput assembly and characterization of PKS pathways in alternative hosts.

Pseudomonas putida Metallothionein: Structural Analysis and Implications of Sustainable Heavy Metal Detoxification in Madinah.

Heavy metals, specifically cadmium (Cd) and lead (Pb), contaminating water bodies of Madinah (Saudi Arabia), is a significant environmental concern that necessitates prompt action. Madinah is exposed to toxic metals from multiple sources, such as tobacco, fresh and canned foods, and industrial activities. This influx of toxic metals presents potential hazards to both human health and the surrounding environment. The aim of this study is to explore the viability of utilizing metallothionein from Pseudomonas putida (P. putida) as a method of bioremediation to mitigate the deleterious effects of pollution attributable to Pb and Cd. The use of various computational approaches, such as physicochemical assessments, structural modeling, molecular docking, and protein-protein interaction investigations, has enabled us to successfully identify the exceptional metal-binding properties that metallothionein displays in P. putida. The identification of specific amino acid residues, namely GLU30 and GLN21, is crucial in understanding their pivotal role in facilitating the coordination of lead and cadmium. In addition, post-translational modifications present opportunities for augmenting the capacity to bind metals, thereby creating possibilities for focused engineering. The intricate web of interactions among proteins serves to emphasize the protein's participation in essential cellular mechanisms, thereby emphasizing its potential contributions to detoxification pathways. The present study establishes a strong basis for forthcoming experimental inquiries, offering potential novel approaches in bioremediation to tackle the issue of heavy metal contamination. Metallothionein from P. putida presents a highly encouraging potential as a viable remedy for environmental remediation, as it is capable of proficiently alleviating the detrimental consequences related to heavy metal pollution.

Potential of algal-based products for the management of potato brown rot disease

BackgroundRalstonia solanacearum causes potato brown rot disease, resulting in lower crop’s production and quality. A sustainable and eco-friendly method for controlling the disease is required. Algae’s bioactive chemicals have shown the potential to enhance plant defenses. For the first time, the efficacy of foliar application of Acanthophora spicifera and Spirulina platensis seaweed extracts, along with the utilization of dried algal biomasses (DABs) of Turbinaria ornata and a mixture of Caulerpa racemosa and Cystoseira myrica (1:1)on potato yield and brown rot suppression were investigated under field conditions. Field experiments were conducted in three locations: Location 1 (Kafr Shukr district, Kaliobeya governorate), Location 2 (Moneira district, Kaliobeya governorate), and Location 3 (Talia district, Minufyia governorate). Locations 1 and 2 were naturally infested with the pathogen, while location 3 was not. The study evaluated potato yield, plant nutritive status and antioxidants, soil available nitrogen-phosphorus-potassium (N-P-K), and organic matter percentage. Additionally, the shift in soil microbial diversity related to R. solanacearum suppression was examined for the most effective treatment.ResultsThe results revealed that seaweed extracts significantly increased potato yield at all locations, which correlated with higher phosphorus absorption, while T. ornate DAB increased potato yield only at location 2, accompanied by noticeable increases in soil nitrogen and plant phosphorus. The mixed DABs of C. racemosa and C. myrica demonstrated greater disease suppression than foliar applications. The disease-suppressive effect of the mixed DABs was accompanied by significant increases in flavonoids and total antioxidant capacity (TAC). Moreover, the application of mixed DABs increased soil bacterial biodiversity, with a higher abundance of oligotrophic marine bacterial species such as Sphingopyxis alaskensis and growth-promoting species like Glutamicibacter arilaitensis, Promicromonospora sp., and Paenarthrobacter nitroguajacolicus in all three locations compared to the untreated control. Klebsiella sp., Pseudomonas putida, and P. brassicacearum abundances were increased by the mixed DABs in Location 1. These species were less abundant in locations 2 and 3, where Streptomyces sp., Bacillus sp., and Sphingobium vermicomposti were prevalent.ConclusionsThe results demonstrated that the used seaweed extracts improved potato yield and phosphorous absorption, while the mixed DABs potentially contributed in disease suppression and improved soil microbial diversity.

Assessing the performance of synthetic co-cultures during the conversion of methane into Poly(3-hydroxybutyrate)

Synthetic co-cultures can enhance pollutant bioconversion performance through synergistic effects among co-existing species. In this study, the potential of Methylocystis hirsuta and Methylocystis parvus to support poly(3-hydroxybutyrate) (PHB) production in co-cultivation with Rhodococcus opacus and Pseudomonas putida under a CH4:O2 atmosphere was assessed batchwise. The metabolic activation of almost all co-cultures studied was faster than that of single strain cultures, bringing higher methane and oxygen consumption rates. Higher PHB yields were achieved when coupling M. hirsuta with R. opacus (63 % w w−1) or with R. opacus and P. putida (64.4 % w w−1) compared to M. hirsuta alone (38.5 % w w−1). Interestingly, the combination of both R. opacus and P. putida with M. parvus reduced PHB accumulation to 42.2 % w w−1 compared to the content observed in M. parvus monocultures (62.2 % w w−1) and M parvus + R. opacus co-cultures (66.6 % w w−1).

Impedance properties of biomass in support of practical mensuration in rain gardens

Impedance microbiology was explored to provide insights into how a sensor that can monitor the growth of bacteria in vegetated bioretention systems (rain gardens) may be designed for in situ, real-time use. The impedance properties of Pseudomonas Putida samples were monitored under AC signals of 100 mV peak-to-peak and sweeping frequencies of 20 Hz–300 kHz, to determine relationships between these properties and biomass in the laboratory. The capacitance of the samples was found to be the most sensitive impedance parameter, with average change in magnitude of 37% due to bacterial growth. For verification, optical density measurements were taken and calibrated by direct hemocytometry counts of similar samples, simultaneously with the impedance testing. The experiments revealed that exponential relationships enable a good estimate of the biomass available in the medium, based on the change in capacitance. The detection range of the proposed system (in the range of tested strain) is approximately ∼9.2 × 106 cells/mL to ∼5 × 108 cells/mL.