Cereal leaf beetle-associated bacteria enhance the survival of their host upon insecticide treatments and respond differently to insecticides with different modes of action.

The cereal leaf beetle (CLB, Oulema melanopus) is one of the major cereal pests. The effect of insecticides belonging to different chemical classes, with different mechanisms of action and the active substances' concentrations on the CLB bacterial microbiome, was investigated. Targeted metagenomic analysis of the V3-V4 regions of the 16S ribosomal gene was used to determine the composition of the CLB bacterial microbiome. Each of the insecticides caused a decrease in the abundance of bacteria of the genus Pantoea, and an increase in the abundance of bacteria of the genus Stenotrophomonas, Acinetobacter, compared to untreated insects. After cypermethrin application, a decrease in the relative abundance of bacteria of the genus Pseudomonas was noted. The dominant bacterial genera in cypermethrin-treated larvae were Lactococcus, Pantoea, while in insects exposed to chlorpyrifos or flonicamid it was Pseudomonas. Insecticide-treated larvae were characterized, on average, by higher biodiversity and richness of bacterial genera, compared to untreated insects. The depletion of CLB-associated bacteria resulted in a decrease in larval survival, especially after cypermethrin and chlorpyrifos treatments. The use of a metagenome-based functional prediction approach revealed a higher predicted function of bacterial acetyl-CoA C-acetyltransferase in flonicamid and chlorpyrifos-treated larvae and tRNA dimethyltransferase in cypermethrin-treated insects than in untreated insects.

Enhancement of herbicolin A production by integrated fermentation optimization and strain engineering in Pantoea agglomerans ZJU23.

BACKGROUND: The lipopeptide herbicolin A (HA) secreted by the biocontrol agent Pantoea agglomerans ZJU23 is a promising antifungal drug to combat fungal pathogens by targeting lipid rafts, both in agricultural and clinical settings. Improvement of HA production would be of great significance in promoting its commercialization. This study aims to enhance the HA production in ZJU23 by combining fermentation optimization and strain engineering. RESULTS: Based on the results in the single-factor experiments, corn steep liquor, temperature and initial pH were identified as the significant affecting factors by the Plackett-Burman design. The fermentation medium and conditions were further optimized using the Box-Behnken response surface method, and the HA production of the wild type strain ZJU23 was improved from ~ 87 mg/mL in King's B medium to ~ 211 mg/mL in HA induction (HAI) medium. A transposon library was constructed in ZJU23 to screen for mutants with higher HA production, and two transcriptional repressors for HA biosynthesis, LrhA and PurR, were identified. Disruption of the LrhA gene led to increased mRNA expression of HA biosynthetic genes, and subsequently improved about twofold HA production. Finally, the HA production reached ~ 471 mg/mL in the ΔLrhA mutant under optimized fermentation conditions, which is about 5.4 times higher than before (~ 87 mg/mL). The bacterial suspension of the ΔLrhA mutant fermented in HAI medium significantly enhanced its biocontrol efficacy against gray mold disease and Fusarium crown rot of wheat, showing equivalent control efficacies as the chemical fungicides used in this study. Furthermore, HA was effective against fungicide resistant Botrytis cinerea. Increased HA production substantially improved the control efficacy against gray mold disease caused by a pyrimethanil resistant strain. CONCLUSIONS: This study reveals that the transcriptional repressor LrhA negatively regulates HA biosynthesis and the defined HAI medium is suitable for HA production. These findings provide an extended basis for large-scale production of HA and promote biofungicide development based on ZJU23 and HA in the future.

In Vitro Antifungal and Antibacterial Effects of Biosynthesized Zinc Oxide Nanoparticles Against Selected Pathogenic and Non-Pathogenic Strains.

We have developed a simple, robust environment-friendly and efficient method for ZnO nanoparticles biosynthesis using Dalbergia sissoo fresh leaf extract. Before using these nanoparticles for antimicrobial assay, a detailed characterization was performed using techniques like Ultraviolet/Visible (UV/Vis) spectroscopy, Particle size analysis (PSA), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), atomic force microscopy (AFM),Transmission electron microscopy (TEM) etc. The average size of biosynthesized ZnO nanoparticles was around 30 nm and they were pure and crystalline by nature. The effectiveness of these biosynthesized nanoparticles were checked against both pathogenic and non-pathogenic microbes. A total of eight bacterial strains-Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Klebsilla pneumoniae, Staphylococcus aureus, Streptococcus entericus, Bacillus cereus, Pantoea cypripedii and three fungal strains-Candida albicans, Aspergilus niger and Aspergilus flavus were studied to have a clear view of the spectrum of ZnO nanoparticles anti-microbial activity. The effectiveness of biosynthesized ZnO nanoparticles against the microbes was found to be better than the standard reference antibiotics used (streptomycin, chloramphenicol and rifampicin). The results seem to be very promising and can be used for some practical applications of ZnO nanoparticles in nearfuture.

Deciphering Microbial Metal Toxicity Responses via Random Bar Code Transposon Site Sequencing and Activity-Based Metabolomics.

To uncover metal toxicity targets and defense mechanisms of the facultative anaerobe Pantoea sp. strain MT58 (MT58), we used a multiomic strategy combining two global techniques, random bar code transposon site sequencing (RB-TnSeq) and activity-based metabolomics. MT58 is a metal-tolerant Oak Ridge Reservation (ORR) environmental isolate that was enriched in the presence of metals at concentrations measured in contaminated groundwater at an ORR nuclear waste site. The effects of three chemically different metals found at elevated concentrations in the ORR contaminated environment were investigated: the cation Al3+, the oxyanion CrO42-, and the oxycation UO22+. Both global techniques were applied using all three metals under both aerobic and anaerobic conditions to elucidate metal interactions mediated through the activity of metabolites and key genes/proteins. These revealed that Al3+ binds intracellular arginine, CrO42- enters the cell through sulfate transporters and oxidizes intracellular reduced thiols, and membrane-bound lipopolysaccharides protect the cell from UO22+ toxicity. In addition, the Tol outer membrane system contributed to the protection of cellular integrity from the toxic effects of all three metals. Likewise, we found evidence of regulation of lipid content in membranes under metal stress. Individually, RB-TnSeq and metabolomics are powerful tools to explore the impact various stresses have on biological systems. Here, we show that together they can be used synergistically to identify the molecular actors and mechanisms of these pertubations to an organism, furthering our understanding of how living systems interact with their environment. IMPORTANCE Studying microbial interactions with their environment can lead to a deeper understanding of biological molecular mechanisms. In this study, two global techniques, RB-TnSeq and activity metabolomics, were successfully used to probe the interactions between a metal-resistant microorganism, Pantoea sp. strain MT58, and metals contaminating a site where the organism can be located. A number of novel metal-microbe interactions were uncovered, including Al3+ toxicity targeting arginine synthesis, which could lead to a deeper understanding of the impact Al3+ contamination has on microbial communities as well as its impact on higher-level organisms, including plants for whom Al3+ contamination is an issue. Using multiomic approaches like the one described here is a way to further our understanding of microbial interactions and their impacts on the environment overall.

Predatory and biocontrol potency of Bdellovibrio bacteriovorus toward phytopathogenic strains of Pantoea sp. and Xanthomonas campestris in the presence of exo-biopolymers: in vitro and in vivo assessments.

Bdellovibrios are predatory bacteria that invade other live Gram-negative bacterial cells for growth and reproduction. They have recently been considered as potential living antibiotics and biocontrol agents. In this study, the predatory activity and biocontrol potency of Bdellovibrio bacteriovorus strain SOIR-1 against Pantoea sp. strain BCCS and Xanthomonas campestris, two exo-biopolymer-producing phytopathogens, was evaluated. Plaque formation assays and lysis analysis in the broth co-cultures were used for the in vitro evaluation of bacteriolytic activity of strain SOIR-1. The in vivo biocontrol potential of strain SOIR-1 was evaluated by pathogenicity tests on the onion bulbs and potato tuber slices. The phytopathogens were also recovered from the infected plant tissues and confirmed using biochemical tests and PCR-based 16S rRNA gene sequence analysis. Typical bdellovibrios plaques were developed on the lawn cultures of Pantoea sp. BCCS and X. campestris. The killing rate of strain SOIR-1 toward Pantoea sp. BCCS and X. campestris was 84.3% and 76.3%, respectively. Exo-biopolymers attenuated the predation efficiency of strain SOIR-1 up to 10.2-18.2% (Pantoea sp. BCCS) and 12.2-17.3% (X. campestris). The strain SOIR-1 significantly reduced rotting symptoms in the onion bulbs caused by Pantoea sp. BCCS (69.0%) and potato tuber slices caused by X. campestris (73.1%). Although more field assessments are necessary, strain SOIR-1 has the preliminary potential as a biocontrol agent against phytopathogenic Pantoea sp. BCCS and X. campestris, especially in postharvest storage. Due to the particular physicochemical properties of evaluated exo-biopolymers, they can be used in the designing encapsulation systems for delivery of bdellovibrios.

Pantoea ananatis carotenoid production confers toxoflavin tolerance and is regulated by Hfq-controlled quorum sensing.

Carotenoids are widely used in functional foods, cosmetics, and health supplements, and their importance and scope of use are continuously expanding. Here, we characterized carotenoid biosynthetic genes of the plant-pathogenic bacterium Pantoea ananatis, which carries a carotenoid biosynthetic gene cluster (including crtE, X, Y, I, B, and Z) on a plasmid. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that the crtEXYIB gene cluster is transcribed as a single transcript and crtZ is independently transcribed in the opposite direction. Using splicing by overlap extension with polymerase chain reaction (SOE by PCR) based on asymmetric amplification, we reassembled crtE-B, crtE-B-I, and crtE-B-I-Y. High-performance liquid chromatography confirmed that Escherichia coli expressing the reassembled crtE-B, crtE-B-I, and crtE-B-I-Y operons produced phytoene, lycopene, and ß-carotene, respectively. We found that the carotenoids conferred tolerance to UV radiation and toxoflavin. Pantoea ananatis shares rice environments with the toxoflavin producer Burkholderia glumae and is considered to be the first reported example of producing and using carotenoids to withstand toxoflavin. We confirmed that carotenoid production by P. ananatis depends on RpoS, which is positively regulated by Hfq/ArcZ and negatively regulated by ClpP, similar to an important regulatory network of E. coli (HfqArcZ →RpoS Í° ClpXP). We also demonstrated that Hfq-controlled quorum signaling de-represses EanR to activate RpoS, thereby initiating carotenoid production. Survival genes such as those responsible for the production of carotenoids of the plant-pathogenic P. ananatis must be expressed promptly to overcome stressful environments and compete with other microorganisms. This mechanism is likely maintained by a brake with excellent performance, such as EanR.

Thiosulfinate Tolerance Is a Virulence Strategy of an Atypical Bacterial Pathogen of Onion.

Unlike most characterized bacterial plant pathogens, the broad-host-range plant pathogen Pantoea ananatis lacks both the virulence-associated type III and type II secretion systems. In the absence of these typical pathogenicity factors, P. ananatis induces necrotic symptoms and extensive cell death in onion tissue dependent on the HiVir proposed secondary metabolite synthesis gene cluster. Onion (Allium. cepa L), garlic (A. sativum L.), and other members of the Allium genus produce volatile antimicrobial thiosulfinates upon cellular damage. However, the roles of endogenous thiosulfinate production in host-bacterial pathogen interactions have not been described. We found a strong correlation between the genetic requirements for P. ananatis to colonize necrotized onion tissue and its capacity for tolerance to the thiosulfinate "allicin" based on the presence of an eleven-gene, plasmid-borne, virulence cluster of sulfur redox genes. We have designated them "alt" genes for allicin tolerance. We show that allicin and onion thiosulfinates restrict bacterial growth with similar kinetics. The alt gene cluster is sufficient to confer allicin tolerance and protects the glutathione pool during allicin treatment. Independent alt genes make partial phenotypic contributions indicating that they function as a collective cohort to manage thiol stress. Our work implicates endogenous onion thiosulfinates produced during cellular damage as major mediators of interactions with bacteria. The P. ananatis-onion pathosystem can be modeled as a chemical arms race of pathogen attack, host chemical counterattack, and pathogen defense.

Phytosynthesis of silver nanoparticles using Mangifera indica flower extract as bioreductant and their broad-spectrum antibacterial activity.

The present study focused on the evaluation of antibacterial property of silver nanoparticles (AgNPs) synthesized using mango flower extract. The morphology of the synthesized AgNPs was observed under transmission electron microscopy and the particles have shown spherical shape in the range of 10-20 nm. X-ray powder diffraction analysis confirmed the crystalline nature of the AgNPs. The atomic percentage of the Ag element in the nanoparticles was about 7.58% which is greater than the other elements present in the sample. The AgNPs showed extensive lethal effect on both Gram-positive (Staphylococcus sp.) and Gram-negative (Klebsiella sp., Pantoea agglomerans, and Rahnella sp.) bacteria. The extensive lethal effect of AgNPs against clinically important pathogens demonstrated that the mango flower mediated AgNPs could be applied as potential antibacterial agent to control the bacterial population in the respective industries.

Identification of Antibacterial Phenolics in Selected Plant Species from Mexican Cloud Forest by Mass Spectrometry Dereplication.

Fifteen plant species from a protected cloud forest (CF) in Veracruz, Mexico, were screened for their in vitro capacity to inhibit the growth of the phytopathogenic bacteria Chryseobacterium sp., Pseudomonas cichorii, Pectobacterium carotovorum and Pantoea stewartii, causal agents of damage to crops like 'chayote', lettuce, potato and corn. As a result, the bioactivity of Turpinia insignis and Leandra cornoides is reported for the first time against Chryseobacterium sp. and P. cichorii. In addition, 24 and 18 compounds not described for these species were dereplicated by an UPLC/MS-MS method, respectively. The identified compounds included simple phenols, hydroxycinnamic acids, flavonoids and coumarins. The antibacterial assay of 12 of them demonstrated the bacteriostatic effect of vanillin, trans-cinnamic acid, scopoletin and umbelliferone against Chryseobacterium sp. These findings confirm for the first time the value of the CF plants from Veracruz as sources of bioactive natural products with antimicrobial properties against phytopathogenic bacteria.

Extracellular DNA enhances the formation and stability of symplasmata in Pantoea agglomerans YS19.

Extracellular DNA (eDNA) is an important polymeric substance that plays essential roles in cell aggregation and nutrient provision for the sessile bacteria. eDNA in bacterial biofilms was extensively studied. Here we found that eDNA also exists in symplasmata, a bacterial cell aggregate, which is different to a biofilm, in the rice enophyte Pantoea agglomerans YS19. We found that exogenous eDNA enhanced the formation and stability of symplasmata significantly, and that, exogenous eDNA also improved the stress resistance and colonization ability of the bacterium on host rice. These results strongly indicate novel roles of the eDNA in Pantoea agglomerans YS19, showing its special relation to the stress-resistance and endophyte-host association of the strain.

Priming with ACC-utilizing bacterium attenuated copper toxicity, improved oxidative stress tolerance, and increased phytoextraction capacity in wheat.

The major challenges for the plants growing in metal-contaminated soils are deficiency of nutrients, biomass reduction, and severe oxidative damages in the presence of heavy metals. In this regard, our aim was to overcome these challenges through the use of efficient microbial strains in metal-polluted soils and to assess its/their physiological and biochemical effects. In the current study, a copper (Cu)-resistant bacterium was isolated from the rhizospheric soil of 'Ziziphus nummularia' and evaluated for its ability to promote the wheat growth under the gradient stress of copper. Based on 16S rRNA gene sequencing, the isolate was identified as Pantoea sp. Among the plant growth promoting tests, the isolate showed the production of indole acetic acid, solubilization of inorganic phosphate, and ACC deaminase activity. Also, the isolate showed resistance to many heavy metals and antibiotics and increased the water-soluble copper in solution. The results of pot studies showed that bacterial application promoted various growth parameters of wheat plants and also enhanced the Cu uptake of wheat from the Cu-amended soil. The results showed that enhancement of Cu stress (100 to 300 mg kg-1) resulted in a decrease in various compatible solutes such as proline, total soluble sugars, and total protein content, and increase in the level of malondialdehyde (MDA), latter of which is the indicator of oxidative stress. Bacterial treatment markedly increased the proline, soluble sugar, total protein content, and decreased the MDA content under Cu stress. In addition, bacterial inoculation significantly alleviated the harmful effect of metal toxicity by decreasing the activation of ROS molecules including superoxide (O2-) and hydrogen peroxide (H2O2). The activation of various antioxidative enzymes such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) was noted following bacterial inoculation under Cu stress. Therefore, the present study demonstrates the potential of the isolate Pantoea sp. ZNP-5 to improve the growth and phytoextraction of metal from the metal-polluted soil through the polyphasic mechanism of action.

Sensitivity of Coriandrum sativum extract on bacterial pathogens isolated from digestive system of rabbits, and its role on in vitro cecal gas production and fermentation.

The present context was aimed to investigate the antibacterial potency of aqueous extract of coriander (Coriandrum sativum L.) leaves against bacterial pathogens isolated from the organs associated with digestive system of rabbit. This study also evaluated the influence of varied doses of aqueous extract of C. sativum (AECS) leaves on in vitro gas production (GP), methane (CH4) production, and some other pivotal fermentation parameters from caecal sample of rabbits. The pathogenic bacteria were isolated from mouth, caecum, and anus of rabbits, and further identified through morphological, biochemical, and molecular tools. The growth inhibitory characteristics of AECS against pathogens were determined using disc diffusion assay. Surprisingly, the result revealed lack of antibacterial potential at tested concentrations. Further, in order to demonstrate the in vitro GP and fermentation parameters in rabbits, four treatments comprising of 0, 0.6, 1.2, and 1.8 mL extract/g dry matter (DM) of AECS were used. Results showed no linear or quadratic effect (P > 0.05) on in vitro GP and CH4 production after the supplementation of AECS in the feeding diet. However, the inclusion of AECS at the concentration of 1.8 mL/g DM exhibited the lowest asymptotic CH4 production and initial delay prior to CH4 production. Similarly, the addition of AECS at 1.8 mL/g DM concentration reduced asymptotic GP as well as CH4 production, and improved fermentation parameters of rabbits when compared with the control and other tested doses. In a nutshell, the tested doses of AECS showed lack of antibacterial trait against the pathogenic bacteria isolated from mouth, caecum, and anus of rabbits. Besides, the AECS exhibited the unique potentiality of reducing GP and improving diversified fermentation parameters in rabbits, thereby suggesting its plausible role as an alternative to commercially available growth promoters in livestock industries.

Transcriptome analysis of silver, palladium, and selenium stresses in Pantoea sp. IMH.

Heavy metal contamination is a significant environmental issue. Using bacteria for removal and reduction of heavy metals is an attractive alternative owing to its low-cost and eco-friendly properties. However, the mechanisms of resistance to and reduction of Ag(I), Pd(II), and Se(IV), especially in the same strain, remain unclear. Here, Pantoea sp. IMH was examed for its reduction of Ag(I), Pd(II), and Se(IV) to nanoparticles (NPs), and the molecular mechanism was investigated by transcriptome analysis. The results revealed that genes encoding binding, transport, catalytic activity, and metabolism were differentially expressed in cells exposed to Ag(I), Pd(II), and Se(IV). The same resistance mechanisms for all metals included multiple stress resistance protein BhsA and glutathione detoxification metabolism. However, zinc transport protein and sulfate metabolism played an important role in the resistance to cationic metals (Ag+ and Pd2+), while the oxalate transporter and arsenic resistance mechanisms were specifically involved in the resistance to and reduction of anion (SeO32-). In addition, Ag(I) was speculated to be reduced to AgNPs by glucose and cytochrome CpxP was involved in Pd(II) reduction. Our results provided new clues on the mechanisms of resistance to and reduction of Ag(I), Pd(II), and Se(IV).

Non-toxic properties of TiO2 and STiO2 nanocomposite PES ultrafiltration membranes for application in membrane-based environmental biotechnology.

In membrane bioreactor (MBR) technology, nanocomposite membrane has a great potential to improve the filtration performance and antifouling. However, antibacterial activity of nanoparticles (NPs) is a significant disadvantage which can be impacted to bacterial growth and microbial community in MBRs. The modified polyethersulfone (PES) ultrafiltration (UF) membranes in the study were prepared by using TiO2 NPs and TiO2 NPs functionalized with sulfonation (STiO2). The antibacterial effect of NPs and non-toxic properties of nanocomposite membranes were examined by using three different Gram-negative bacterial species isolated from a local full scale membrane bioreactor treating municipal wastewater (Escherichia coli, Pantoea agglomerans, and Pseudomonas graminis). Results are revealed that the TiO2 and STiO2 NPs have 60% of antibacterial activity based on disc diffusion, viability tests, and TEM analysis. However, the PES-TiO2 and PES-STiO2 nanocomposite UF membranes showed significantly lower antibacterial activity (<95%, significance at p < 0.0001), indicating innocuous to bacterial growth. This study highlights that the PES-TiO2 and PES-STiO2 nanocomposite membrane is more sustainable than PES membrane and promising materials for MBRs, by taking advantage of non-toxic properties to bacterial growth.

Impact of modified diamond-like carbon coatings on the spatial organization and disinfection of mixed-biofilms composed of Escherichia coli and Pantoea agglomerans industrial isolates.

This work investigated the effects of diamond-like carbon (DLC) coatings on the architecture and biocide reactivity of dual-species biofilms mimicking food processing contaminants. Biofilms were grown using industrial isolates of Escherichia coli and Pantoea agglomerans on bare stainless steel (SST) and on two DLC surface coatings (a-C:H:Si:O designated by SICON® and a-C:H:Si designated by SICAN) in order to evaluate their antifouling activities. Quantification and spatial organization in single- and dual-species biofilms were examined by confocal laser scanning microscopy (CLSM) using a strain specific labelling procedure. Those assays revealed that the E. coli isolate exhibited a higher adhesion to the modified surfaces and a decreased susceptibility to disinfectant in presence of P. agglomerans than alone in axenic culture. While SICON® reduced the short-term growth of E. coli in axenic conditions, both DLC surfaces increased the E. coli colonization in presence of P. agglomerans. However, both modified surfaces triggered a significantly higher log reduction of E. coli cells within mixed-species biofilms, thus the use of SICON® and SICAN surfaces may be a good approach to facilitate the disinfection process in critical areas of food processing plants. This study presents a new illustration of the importance of interspecies interactions in surface-associated community functions, and of the need to evaluate the effectiveness of hygienic strategies with relevant multi-species consortia.

Antimicrobial resistance and molecular characterization of Pantoea agglomerans isolated from rainbow trout (Oncorhynchus mykiss) fry.

Aquaculture has become an important candidate as an animal protein source through its growth over the last decade. Based upon a report from the Food and Agriculture Organization of the United Nations, it is the fastest growing sector of the food industry, yet the pathogenicity of many biological agents involved in aquaculture is still unknown. In this study, we isolated Pantoe agglomerans from diseased rainbow trout on several occasions and also attempted to determine their phenotypic and genotypic characteristics, including antimicrobial resistance, of four bacterial isolates. In the present study, P. agglomerans was isolated from diseased rainbow trout as a pathogenic agent. The identification of the P. agglomerans isolates from the rainbow trout was performed through biochemical tests and 16S rRNA sequence analysis. These isolates were predominately biochemically homogeneous, although some features were different, such as seen in methyl-red, mannose and lipase activity tests. All four studied isolates were identified as 99% identical to P. agglomerans based on sequence analysis. The isolates were compared through a phylogenetic analysis with P. agglomerans sequences recovered from 16 other countries and accessed from the GenBank database. All isolates in our study were at least 98.2% similar to sequences from GenBank. Furthermore, the phenotypic antimicrobial susceptibility of the isolates in this study was analyzed through both disc diffusion and broth micro dilution minimum inhibitory concentration (MIC) tests. Although there were some differences between two phenotypic antimicrobial tests, all studied isolates were found susceptible to different antimicrobials. In addition genotypic antimicrobial resistance characteristics were assessed by the presence of antimicrobial resistance genes (ARGs), in which qnrS and sul2 were detected for the first time in P. agglomerans.

Heavy Metal Stress and Its Consequences on Exopolysaccharide (EPS)-Producing Pantoea agglomerans.

Currently, the heavy metal pollution is of grave concern, and the part of microorganism for metal bioremediation should take into account as an efficient and economic strategy. On this framework, the heavy metal stress consequences on exopolysaccharide (EPS)-producing agricultural isolate, Pantoea agglomerans, were studied. The EPS production is a protective response to stress to survive and grow in the metal-contaminated environment. P. agglomerans show tolerance and mucoid growth in the presence of heavy metals, i.e., mercury, copper, silver, arsenic, lead, chromium, and cadmium. EDX first confirmed the metal accumulation and further, FTIR determined the functional groups involved in metal binding. The ICP-AES identified the location of cell-bound and intracellular metal accumulation. Metal deposition on cell surface has released more Ca2+. The effect on bacterial morphology investigated with SEM and TEM revealed the sites of metal accumulation, as well as possible structural changes. Each heavy metal caused distinct change and accumulated on cell-bound EPS with some intracellular deposits. The metal stress caused a decrease in total protein content and increased in total carbohydrate with a boost in EPS. Thus, the performance of P. agglomerans under metal stress indicated a potential candidate for metal bioremediation. Graphical Abstract ᅟ.

Clinical and microbiological characteristics of Pantoea agglomerans infection in children.

Pantoea agglomerans is an environmental Gram-negative bacterium that rarely is responsible for the infections in humans but it is often a causative factor of a number of occupational diseases. This study evaluated the clinical and microbiological characteristics and pathogenicity of P. agglomerans in children. We retrospectively reviewed microbiological test results for all children (1 month old to 18 years old) who were admitted to our pediatric hospital between January 2000 to June 2015 and had positive clinical specimen cultures for P. agglomerans. Isolates were identified using conventional tests and the BBL Crystal E/NF ID or MALDI-TOF MS systems. Antibiotic susceptibilities were evaluated using the Kirby-Bauer disc diffusion method. We identified fifteen positive cultures from 14 patients with confirmed infections. The positive specimens included pus, urine, tracheal aspirate, blood, and central venous line samples that yielded P. agglomerans. The median patient age was 8.8 years (range: 1.5 months to 16.5 years), and all patients had underlying comorbidities. Five patients had medical devices, and two devices were removed. The most common P. agglomerans infections involved wound infections (35.7%), pneumonia (21.4%), and urinary tract infections (21.4%). Three patients had concomitant infections (Enterococcus faecium, Pseudomonas aeruginosa, and Aspergillus fumigatus). Five patients had anemia. Three patients (21.4%) died, and all three had carbapenem-resistant P. agglomerans that was detected after the first week of hospitalization; two cases involved pneumonia, which was ineffectively treated. P. agglomerans infections may be life-threatening, especially in young patients with pneumonia. Hospital-acquired P. agglomerans may have different pathogenicity and clinical features, compared to community-acquired P. agglomerans, although further studies are needed to understand the drug-resistance patterns in this bacterium.

Efficient antibacterial nanosponges based on ZnO nanoparticles and doxycycline.

Bacterial soft rot is responsible for the loss of about 25% of worldwide production in vegetables and fruits. Efforts have been made to develop an effective nanosponge with the capacity to load and release antibacterial drugs to protect plants. Based on the potential of the ZnO nanoparticles (ZnO-NPs) to achieve this goal, this study synthesized NP via the sol-gel and hydrothermal methods by controlling native defects, such as oxygen vacancies, using thermal treatments and reduced atmospheres. To characterize the ZnO NPs, X-ray diffraction (XRD), Raman spectroscopy, transmission electron microscopy (TEM), optical spectroscopy, electron paramagnetic resonance (EPR), Zeta Potential measurements and surface area with the Brunauer-Emmett-Teller (BET) method were used. The photophysical and photochemical properties via spin trapping method aligned with EPR using UVA light showed a greater formation of electron-hole pairs and hydroxyl radicals for the reduced ZnO NPs when compared with the oxidized ones. Additionally, we found that reduced ZnO-NPs have high effectively against Escherichia coli, Erwinia carotovora and Pantoea sp. bacteria using the photocatalytic effect in the UV range. Moreover, ZnO-NPs loaded with DOX release profile enables the release of DOX within 46days, where 25% was released during the first 10h followed by a second delivery phase with an interesting short-term efficacy (<1day) against E. carotovora and Pantoea sp. Bacteria. For the first time, it was demonstrated that ZnO-NPs and ZnO-NPs loaded with DOX have efficient UV photocatalytic activities against bacterial soft rot infections.

Indole enhances the survival of Pantoea ananatis YJ76 in face of starvation conditions.

Pantoea ananatis YJ76 is an indole-producing predominant diazotrophic endophyte isolated from rice having multiple growth-promoting effects on host plant. As a decomposition metabolite of L-tryptophan (L-Trp), indole is confirmed to regulate various physiological processes of bacteria. In this research, we found that indole significantly improves the survival of YJ76 in face of starvation conditions and the promoting effect is related to the glycogen accumulation promoted by indole, which is much more significant in the middle decline phase than in other growth phases. Since carbon storage regulator CsrA is a key inhibiting factor on the storage of glycogen in bacteria, we explored the relation between indole-enhanced glycogen accumulation and csrA expression and found that there is a positive correlation between indole-enhanced glycogen accumulation and the indole-inhibited csrA expression in YJ76, which implies the potential relation between CsrA regulation and indole regulatory pathway.