Extracellular Compounds Research Articles

Unknown Extracellular and Bioactive Metabolites of the Genus Alexandrium: A Review of Overlooked Toxins.

Various species of Alexandrium can produce a number of bioactive compounds, e.g., paralytic shellfish toxins (PSTs), spirolides, gymnodimines, goniodomins, and also uncharacterised bioactive extracellular compounds (BECs). The latter metabolites are released into the environment and affect a large range of organisms (from protists to fishes and mammalian cell lines). These compounds mediate allelochemical interactions, have anti-grazing and anti-parasitic activities, and have a potentially strong structuring role for the dynamic of Alexandrium blooms. In many studies evaluating the effects of Alexandrium on marine organisms, only the classical toxins were reported and the involvement of BECs was not considered. A lack of information on the presence/absence of BECs in experimental strains is likely the cause of contrasting results in the literature that render impossible a distinction between PSTs and BECs effects. We review the knowledge on Alexandrium BEC, (i.e., producing species, target cells, physiological effects, detection methods and molecular candidates). Overall, we highlight the need to identify the nature of Alexandrium BECs and urge further research on the chemical interactions according to their ecological importance in the planktonic chemical warfare and due to their potential collateral damage to a wide range of organisms.

Secreted acid phosphatases maintain replicative lifespan via inositol polyphosphate metabolism in budding yeast.

Secreted acid phosphatases (APases) dephosphorylate extracellular organic phosphate compounds to supply inorganic phosphate (Pi) to maintain cellular functions. Here, we show that APases are necessary to maintain a normal replicative lifespan in Saccharomyces cerevisiae. Deletion of all four APase genes shortened the lifespan in yeast strains on synthetic media (irrespective of the concentrations of Pi in the media), but it did not affect the intracellular ortho- and polyphosphate levels. Deletion of inositol-pentakisphosphate 2-kinase (IPK1), which encodes inositol-pentakisphosphate 2-kinase, restored the lifespan in APase-null mutants, and IPK1 overexpression shortened the lifespan in wild-type strains. Overexpression of inositol hexakisphosphate (IP6 ) and heptakisphosphate kinases, KCS1 and VIP1, recovered the lifespan in APase-null mutants. Thus, yeast APases modulate the replicative lifespan, probably through dephosphorylation of intracellular IP6 .

Host genotype structures the microbiome of a globally dispersed marine phytoplankton

Phytoplankton support complex bacterial microbiomes that rely on phytoplankton-derived extracellular compounds and perform functions necessary for algal growth. Recent work has revealed sophisticated interactions and exchanges of molecules between specific phytoplankton-bacteria pairs, but the role of host genotype in regulating those interactions is unknown. Here, we show how phytoplankton microbiomes are shaped by intraspecific genetic variation in the host using global environmental isolates of the model phytoplankton host Thalassiosira rotula and a laboratory common garden experiment. A set of 81 environmental T. rotula genotypes from three ocean basins and eight genetically distinct populations did not reveal a core microbiome. While no single bacterial phylotype was shared across all genotypes, we found strong genotypic influence of T. rotula, with microbiomes associating more strongly with host genetic population than with environmental factors. The microbiome association with host genetic population persisted across different ocean basins, suggesting that microbiomes may be associated with host populations for decades. To isolate the impact of host genotype on microbiomes, a common garden experiment using eight genotypes from three distinct host populations again found that host genotype influenced microbial community composition, suggesting that a process we describe as genotypic filtering, analogous to environmental filtering, shapes phytoplankton microbiomes. In both the environmental and laboratory studies, microbiome variation between genotypes suggests that other factors influenced microbiome composition but did not swamp the dominant signal of host genetic background. The long-term association of microbiomes with specific host genotypes reveals a possible mechanism explaining the evolution and maintenance of complex phytoplankton-bacteria chemical exchanges.

Assessing the accumulation efficiency of various microbial carbon components in soils of different minerals

Heterotrophic soil microbes are increasingly recognized as a key mediator transforming labile organic carbon (OC) into relatively stable soil carbon (C) in the form of microbial necromass (dead cells) and extracellular compounds associated with minerals. However, the accumulation of microbial necromass relative to labile OC consumption and its regulating factors remain poorly understood, although it has vital implications for soil C sequestration and modeling. Here by mimicking microbial C accumulation in constructed model soils using fructose as the sole OC substrate, we present a benchmark comparison of microbial C accrual versus OC mineralization under declining substrate availability and mineralogy. By quantifying various microbial components including biomass (living cells; indicated by phospholipid fatty acids), necromass (indicated by amino sugars) and total microbial C (including biomass, necromass and extracellular compounds; estimated as the difference between added and residual substrate C minus respiration) in a simple soil system, we compare microbial metabolic quotient (qCO2; i.e., microbial respiration rate per unit of biomass), amino sugar accumulation efficiency (AAE; i.e., ratio of amino sugars to respiration) and microbial C accumulation efficiency (mCAE; i.e., ratio of total microbial C to total microbial C and respiration), and assess their regulating factors. We find that while clay (bentonite) promotes microbial respiration, it enhances the rate as well as efficiency of amino sugar accumulation without affecting qCO2 or mCAE. On the contrary, ferrihydrite increases qCO2 and decreases AAE but promotes labile OC preservation via inhibiting microbial growth in the alkaline model soil. Hence, amino sugar accrual is more efficient in clay-rich model soils while labile OC is less consumed in model soils containing iron (hydr)oxide. Furthermore, while mCAE was correlated with qCO2 in all but the model soil with 6% clay, AAE was correlated with qCO2 only in model soils with 12% clay when the mineral treatments were considered separately. Collectively, our findings suggest that mCAE and AAE heavily depend on mechanisms preserving microbial C components but not solely on the metabolic efficiency and is mediated by soil mineral content as well as composition. Parameters considering microbial C preservation such as mCAE or AAE warrant further study for modelling and managing the formation of microbial derived stable soil OC.

The antioxidant activity of Lactic acid bacteria and probiotics: a review

Probiotics and Lactic Acid Bacteria play important roles such as the production of antimicrobial compounds and other metabolites. So they have positive effects on human health. When reactive oxygen species generated in excess or cellular defenses are deficient, biomolecules can be damaged by the oxidative stress process. Various studies have shown that the best way to protect the human body from the effects of oxidation reactions is to avoid them, which can be accomplished by using antioxidants. Due to the damages of synthetic antioxidants, their usage has been discussed. Nowadays natural antioxidants derived from bio-resources have recently gained a lot of attention as a potential replacement for synthetic antioxidants. Probiotic bacteria are thought to defend against oxidative stress by restoring the gut microbiota, according to hypothesis of some scientists. This type of microorganisms indicated their antioxidant activity by producing and increasing antioxidant enzymes, production of secondary metabolites, small hydrolyzed peptides in food, resistance to the presence of hydrogen peroxide, and production of intracellular and extracellular compounds such as Exopolysaccharides. Also, they have shown their positive effect on in vivo models. In conclusion, according to the results of studies, lactic acid bacteria and probiotics are significant sources of natural antioxidants. Therefore, they have important research value and market development potential. Also, it should be noted that the mechanism of antioxidant activity of this group of microorganisms has not been fully investigated, this requires further research.

The effect of cartilage decellularized extracellular matrix-chitosan compound on treating knee osteoarthritis in rats.

Knee osteoarthritis (KOA) refers to a common disease in orthopaedics, whereas effective treatments have been rarely developed. As indicated from existing studies, chondrocyte death, extracellular matrix degradation and subchondral bone injury are recognized as the pathological basis of KOA. The present study aimed to determine the therapeutic effect of decellularized extracellular matrix-chitosan (dECM-CS) compound on KOA. In this study, rat knee cartilage was decellularized, and a satisfactory decellularized extracellular matrix was developed. As suggested from the in vitro experiments, the rat chondrocytes co-cultured with allogeneic dECM grew effectively. According to the results of the alamar blue detection, dECM did not adversely affect the viability of rat chondrocytes, and dECM could up-regulate the genes related to the cartilage synthesis and metabolism. As reported from the animal experiments, dECM-CS compound could protect cartilage, alleviate knee joint pain in rats, significantly delay the progress of KOA in rats, and achieve high drug safety. In brief, dECM-CS compound shows a good therapeutic effect on KOA.

Honey bee-associated bacteria as producers of bioactive compounds for protecting hives. A biosynthetic gene-based approach

Honey bee-associated bacteria are a source of natural compounds of interest for controlling hive decline which is threatening bee health globally. Genes involved in the biosynthesis of a series of extracellular compounds released by bacteria living on the external surface of honey bees were investigated. A biosynthetic gene-based approach was adopted by developing a battery of primers to target the genes involved in the biosynthesis of four groups of bioactive compounds (pyrrolizidine alkaloids, surfactin, 2-heptanone and helveticin J). The primers were tested on 51 bacterial isolates belonging to Bacillus thuringiensis, Acetobacteraceae bacterium, Bifidobacterium asteroides and Apilactobacillus kunkeei. The developed primers led to species-specific detection and characterization of the functional genes involved in the production of three out of four groups of compounds selected for this study. The findings suggest that microbial populations inhabiting apiaries harbor genes involved in the biosynthesis of metabolites linked to the reduction of important honey bee pathogens such as Varroa destructor, Paenibacillus larvae and Nosema ceranae. The gene-based approach adopted for evaluating the biosynthetic potential of bioactive compounds in hives is promising for investigating further compounds for low input control strategies of bee enemies.

Overview of the Role of Rhizobacteria in Plant Salt Stress Tolerance

Salinity is one of the main causes of abiotic stress in plants, resulting in negative effects on crop growth and yield, especially in arid and semi-arid regions. The effects of salinity on plant growth mainly generate osmotic stress, ion toxicity, nutrient deficiency, and oxidative stress. Traditional approaches for the development of salt-tolerant crops are expensive and time-consuming, as well as not always being easy to implement. Thus, the use of plant growth-promoting bacteria (PGPB) has been reported as a sustainable and cost-effective alternative to enhance plant tolerance to salt stress. In this sense, this review aims to understand the mechanisms by which PGPB help plants to alleviate saline stress, including: (i) changes in the plant hormonal balance; (ii) release of extracellular compounds acting as chemical signals for the plant or enhancing soil conditions for plant development; (iii) regulation of the internal ionic content of the plant; or iv) aiding in the synthesis of osmoprotectant compounds (which reduce osmotic stress). The potential provided by PGPB is therefore an invaluable resource for improving plant tolerance to salinity, thereby facilitating an increase in global food production and unravelling prospects for sustainable agricultural productivity.

Sterculic Acid Alters Adhesion Molecules Expression and Extracellular Matrix Compounds to Regulate Migration of Lung Cancer Cells.

Simple SummarySterculic acid (SA) is a naturally occurring lipid with SCD1 inhibitory activity, but it also modifies many other pathways and underlying gene expression. SCD upregulation has been associated with tumor aggressiveness and progression. Effects of SA treatment over extracellular matrix compounds and adhesion molecule expression have not been described in cancer cells up to now. Our results show that SA induces cell death at high dose, but we also observed that lower concentrations of SA treatments also reduce cell adhesion-migration and modify integrins and extracellular matrix compounds expression.Sterculic acid (SA) is a cyclopropenoid fatty acid isolated from Sterculia foetida seeds. This molecule is a well-known inhibitor of SCD1 enzyme, also known as ∆9-desaturase, which main function is related to lipid metabolism. However, recent studies have demonstrated that it also modifies many other pathways and the underlying gene expression. SCD overexpression, or up-regulated activity, has been associated with tumor aggressiveness and poor prognosis in many cancer types. Scd1 down-regulation, with different inhibitors or molecular strategies, reduces tumor cell survival and cell proliferation, as well as the chemoresistance associated with cancer stem cell presence. However, SA effects over cancer cell migration and extracellular matrix or adhesion molecules have not been described in cancer cells up to now. We used different migration assays and qPCR gene expression analysis to evaluate the effects of SA treatment in cancer cells. The results reveal that SA induces tumoral cell death at high doses, but we also observed that lower SA-treatments induce cell adhesion-migration capacity reduction as a result of modifications in the expression of genes related to integrins and extracellular matrix compounds. Overall, the functional and transcriptomic findings suggest that SA could represent a new inhibitor activity of epithelial to mesenchymal transition.

Antimicrobial activity of endophytic fungi isolated from different plant parts of Curcuma mangga Valeton and Zijp

The endophytic fungi isolated from different plant parts including leaf, stem and rhizome of Curcuma mangga were screened for antimicrobial activity by employing agar plug diffusion assay and disc diffusion assay for primary screening and secondary screening, respectively. A total of 127 endophytic fungi that were successfully isolated from various plant parts were cultured to examine their antimicrobial activities. Qualitative screening using agar plug diffusion assay revealed that 118 isolates (92.9%) showed antimicrobial activity against at least on one test microorganisms and suggested that the rhizome part exhibited the highest percentage of antiyeast (58.3%) and antifungal (91.7%) activities compared to leaf and stem parts. Quantitative screening using disc diffusion assay indicated that ethyl acetate extract from fermentative broth (extracellular compound) demonstrated better antimicrobial activity compared to methanol extract derived from fungal biomass (intracellular compound) against all the four classes of pathogenic microorganisms tested (Gram-positive bacteria, Gram-negative bacteria, yeast and fungi). The future of endophytic fungus study is very promising as it possesses hidden potential to be developed as natural antimicrobial agent.

Simultaneous removal of hydrogen sulfide, phosphate and emerging organic contaminants, and improvement of sludge dewaterability by oxidant dosing in sulfide-iron-laden sludge

Liquid sludge often contains odorous and toxic hydrogen sulfide and high levels of FeII compounds (e.g., iron sulfides), due to the extensive use of iron salts for hydrogen sulfide control in sewers and for enhanced primary treatment and phosphate removal in wastewater treatment plants. We proposed and verified that dosing appropriate chemical oxidants in the sulfide-iron-laden sludge can be a simple and cost-effective strategy to remove hydrogen sulfide, phosphate, and emerging organic contaminants, and to improve sludge dewaterability simultaneously. Among the seven oxidants investigated, H2O2, ClO2 and NaClO2 were the more cost-effective oxidants than others to control hydrogen sulfide release from the liquid sludge. Dosing these three oxidants also improved sludge dewaterability and removed dissolved phosphate from the liquid sludge, with H2O2 performing the best. Hydrogen sulfide was removed via both direct oxidation by the dosed oxidants and indirect oxidation by the FeIII that was in-situ formed from oxidation of the FeII compounds in the sludge. The in-situ formed FeIII also precipitated/adsorbed the soluble phosphate into the solid form (FePO4). Fenton-like reactions occurred between H2O2 and the FeII compounds in the sludge, and hydroxyl radicals (HO•) were generated. HO• oxidized hydrogen sulfide, destructed refractory organic emerging contaminants and sludge extracellular polymeric compounds (EPSs), and improved the sludge dewaterability. The formation of HO• can be enhanced by hydrogen sulfide and the sludge EPSs present in the sludge through providing more available FeII for the Fenton-like reactions. This study demonstrates the importance of selecting and dosing suitable oxidants to the sulfide-iron-laden sludge with due consideration for the multiple benefits in engineering practices. The same principles may be also used in formulating a dual oxidant-iron strategy to treat sulfide-iron-laden sewage, sludge, and sediments for simultaneous abatement of various pollutants.

In Vitro Effects of Paralytic Shellfish Toxins and Lytic Extracellular Compounds Produced by Alexandrium Strains on Hemocyte Integrity and Function in Mytilus edulis.

Harmful effects caused by the exposure to paralytic shellfish toxins (PSTs) and bioactive extracellular compounds (BECs) on bivalves are frequently difficult to attribute to one or the other compound group. We evaluate and compare the distinct effects of PSTs extracted from Alexandrium catenella (Alex5) cells and extracellular lytic compounds (LCs) produced by A. tamarense (NX-57-08) on Mytilus edulis hemocytes. We used a 4 h dose–response in vitro approach and analyzed how these effects correlate with those observed in a previous in vivo feeding assay. Both bioactive compounds caused moderated cell death (10–15%), being dose-dependent for PST-exposed hemocytes. PSTs stimulated phagocytic activity at low doses, with a moderate incidence in lysosomal damage (30–50%) at all tested doses. LCs caused a dose-dependent impairment of phagocytic activity (up to 80%) and damage to lysosomal membranes (up to 90%). PSTs and LCs suppressed cellular ROS production and scavenged H2O2 in in vitro assays. Neither PSTs nor LCs affected the mitochondrial membrane potential in hemocytes. In vitro effects of PST extracts on M. edulis hemocytes were consistent with our previous study on in vivo exposure to PST-producing algae, while for LCs, in vivo and in vitro results were not as consistent.

Mast cell chymase affects the functional properties of primary human airway fibroblasts: Implications for asthma

Mast cells (MCs) have a profound impact on allergic asthma. Under such conditions, MCs undergo degranulation, resulting in the release of exceptionally large amounts of MC-restricted proteases. However, the role of these proteases in asthma is only partially understood. We sought to test our hypothesis that MC proteases can influence the functionality of human lung fibroblasts (HLFs). Primary HLFs were treated with MC chymase or tryptase, followed by assessment of parameters related to fibroblast function. HLFs underwent major morphologic changes in response to chymase, showing signs of cellular contraction, but were refractory to tryptase. However, no effects of chymase on HLF viability or proliferation were seen. Chymase, but not tryptase, had a major impact on the output of extracellular matrix-associated compounds from the HLFs, including degradation of fibronectin and collagen-1, and activation of pro-matrix metalloprotease 2. Further, chymase induced the release of various chemotactic factors from HLFs. In line with this, conditioned medium from chymase-treated HLFs showed chemotactic activity on neutrophils. Transcriptome analysis revealed that chymase induced a proinflammatory gene transcription profile in HLFs, whereas tryptase had minimal effects. Chymase, but not tryptase, has a major impact on the phenotype of primary airway fibroblasts by modifying their output of extracellular matrix components and by inducing a proinflammatory phenotype.

Bacterial Extracellular Polymers: A Review

Prokaryotic microbial cells especially bacteria are highly emphases for their exopolysaccharides (EPS) production. EPS are the higher molecular weight natural extracellular compounds observe at the surface of the bacterial cells. Nowadays bacterial EPS represent rapidly emerging as new and industrially important biomaterials because it having tremendous physical and chemical properties with novel functionality. Due to its industrial demand as well as research studies the different extraction processes have been discovered to remove the EPS from the microbial biofilm. The novelties of EPS are also based on the microbial habitat conditions such as higher temperature, lower temperature, acidic, alkaliphilic, saline, etc. Based on its chemical structure they can be homopolysaccharide or heteropolysaccharide. EPSs have a wide range of applications in various industries such as food, textile, pharmaceutical, heavy metal recovery, agriculture, etc. So, this review focus on the understanding of the structure, different extraction processes, biosynthesis and genetic engineering of EPS as well as their desirable biotechnological applications.

Production of Biopolymer from Bacteria - A Review

Polyhydroxyalkanoates (PHA) producers have been found in a variety of ecological niche’s that are naturally or unintentionally exposed to high organic matter or growth limiting substances such as dairy wastes, hydrocarbon contaminated sites, pulp and paper mill wastes, agricultural wastes, activated sludges of treatment plants, rhizosphere, and industrial effluents. Few of them also create extracellular byproducts such as rhamnolipids, extracellular polymeric compounds, and biohydrogen gas. These microbes can use waste materials of various origin as substrates while producing valuable bioproducts such as PHB. As a result, these microbes are industrially important candidates for production; Implementation of an integrated system to separate their by-products (intracellular and extracellular) could be an economical method. In this study, we reviewed several microorganisms that live in diverse environmental situations and are stimulated to collect carbon as polyhydroxyalkanoates granules, as well as variables that influence their production and composition. Ultimately, the current cost of bioplastic manufacture from stored PHA granules can be decreased by investigating capabilities such as dual generation of microorganisms and utilization of wastes as renewable substrate under optimal growth conditions in either a batch or continuous process.

The toxic dinoflagellate Alexandrium minutum affects oyster gamete health and fertilization potential

Dinoflagellates from the globally distributed genus Alexandrium are known to produce both paralytic shellfish toxins (PST) and uncharacterized bioactive extracellular compounds (BEC) with allelopathic, ichthyotoxic, hemolytic and cytotoxic activities. In France, blooms of Alexandrium minutum appear generally during the spawning period of most bivalves. These blooms could therefore alter gametes and/or larval development of bivalves, causing severe issues for ecologically and economically important species, such as the Pacific oyster Crassostrea (=Magallana) gigas. The aim of this work was to test the effects of three strains of A. minutum producing either only PST, only BEC, or both PST and BEC upon oyster gametes, and potential consequences on fertilization success. Oocytes and spermatozoa were exposed in vitro for 2 h to a range of environmentally realistic A. minutum concentrations (10–2.5 × 104 cells mL−1). Following exposure, gamete viability and reactive oxygen species (ROS) production were assessed by flow cytometry, spermatozoa motility and fertilization capacities of both spermatozoa and oocytes were analysed by microscopy. Viability and fertilization capacity of spermatozoa and oocytes were drastically reduced following exposure to 2.5 × 104 cells mL−1 of A. minutum. The BEC-producing strain was the most potent strain decreasing spermatozoa motility, increasing ROS production of oocytes, and decreasing fertilization, from the concentration of 2.5 × 103 cells mL−1. This study highlights the significant cellular toxicity of the BEC produced by A. minutum on oyster gametes. Physical contact between gametes and motile thecate A. minutum cells may also contribute to alter oyster gamete integrity. These results suggest that oyster gametes exposure to A. minutum blooms could affect oyster fertility and reproduction success.

Edible coatings for strawberry based on extracellular compounds of Humphreya coffeata

Edible coatings (EC) are a biodegradable alternative for food preservation. In this work EC based on extracellular compounds of Humphreya coffeata and pectin (PHC) were elaborated and characterized through testing on strawberries. Two PHC types were obtained (PHC1, which settled, and PHC2, which floated during the first separation). The conservation period of the strawberry and the phenol content and antioxidant activity in strawberries with the EC were determined. Four EC were prepared (EC1, EC2, EC3, and EC4), in which EC1 did not contain any PHC, EC2 contained PHC1, EC3 contained PHC2, and EC4 contained both PHCs. Three EC types (EC2, EC3, and EC4) delayed the decay of strawberries approximately 90% at 20 °C during the first 10 days of evaluation, and at 4 °C the effect was between 75 and 90% after 25 days of experimentation. However, EC3 and EC4, both of which contained PHC2, presented the best results. The content of total phenols (approximately 55 mg (gallic acid equivalent (GAE)/mL) and the free radical scavenging activity were 35% with ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 75% with DPPH (2,2-diphenyl-1-picrylhydrazyl) in strawberries with EC. Thus, EC are an alternative for preserving strawberries without losing their antioxidant capacity.

Exendin-4 differentially modulates essential functions of human dermal fibroblasts under normoglycemic and hyperglycemic conditions.

Evidence that exendin-4, a glucagon-like peptide-1 analog, might be used to treat poorly healing wounds under diabetic and nondiabetic conditions has gained increasing interest. Little is known, however, about the effects of the drug on the production by dermal fibroblasts of key extracellular matrix and regulatory compounds. Therefore, we used human skin fibroblasts cultured in normo- (1 g/l = 5.6 mmol/l glucose) or hyperglycemic (4.5 g/l = 25 mmol/l glucose) culture medium to test the effects of exendin-4 (0 - 100 nmol/l) on fibroblast functions crucial for the wound healing process. Exendin-4 increased the proliferative and metabolic activities, as measured by the BrdU (bromodeoxyuridine) and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assays, respectively, of fibroblasts cultured in normoglycemic medium. Under hyperglycemic conditions, the drug had no effect on proliferation and reduced metabolic fibroblast activity. Exendin-4 decreased metalloproteinase-9 (MMP-9) secretion in the normoglycemic milieu only and increased tissue inhibitor of metalloproteinase-1 (TIMP-1) concentration in fibroblast colonies under both normo- and hyperglycemic experimental conditions. Exendin-4 increased the fibroblast growth factor-1 (FGF-1) concentration in cell colonies maintained in the normoglycemic milieu but decreased FGF-1 release when fibroblasts were grown in hyperglycemic medium. High glucose caused lactic dehydrogenase (LDH) leakage when compared with normoglycemic conditions, and exendin-4 was not able to prevent this effect, although it reduced LDH release from fibroblasts cultured in normoglycemic medium. Finally, exendin-4 increased glycosaminoglycan (GAG) content under both experimental conditions. Our results indicate that exendin-4 effects on the production of the extracellular matrix and regulatory proteins differ in human skin fibroblasts exposed to either normal or high glucose. In general, the beneficial effects of the drug, which may be important for the improvement of wound healing, are more pronounced under normoglycemic conditions, thus indicating that hyperglycemia attenuates the positive effects of exendin-4 on fibroblasts.

Characterization of the biocontrol activity of three bacterial isolates against the phytopathogen Erwinia amylovora.

Antibiotics are sprayed on apple and pear orchards to control, among other pathogens, the bacterium Erwinia amylovora, the causative agent of fire blight. As with many other pathogens, we observe the emergence of antibiotic‐resistant strains of E. amylovora. Consequently, growers are looking for alternative solutions to combat fire blight. To find alternatives to antibiotics against this pathogen, we have previously isolated three bacterial strains with antagonistic and extracellular activity against E. amylovora, both in vitro and in planta, corresponding to three different bacterial genera: Here, we identified the inhibitory mode of action of each of the three isolates against E. amylovora. Isolate Bacillus amyloliquefaciens subsp. plantarum (now B. velezensis) FL50S produces several secondary metabolites including surfactins, iturins, and fengycins. Specifically, we identified oxydifficidin as the most active against E. amylovora S435. Pseudomonas poae FL10F produces an active extracellular compound against E. amylovora S435 that can be attributed to white‐line‐inducing principle (WLIP), a cyclic lipopeptide belonging to the viscosin subfamily (massetolide E, F, L, or viscosin). Pantoea agglomerans NY60 has a direct cell‐to‐cell antagonistic effect against E. amylovora S435. By screening mutants of this strain generated by random transposon insertion with decreased antagonist activity against strain S435, we identified several defective transposants. Of particular interest was a mutant in a gene coding for a Major Facilitator Superfamily (MFS) transporter corresponding to a transmembrane protein predicted to be involved in the extracytoplasmic localization of griseoluteic acid, an intermediate in the biosynthesis of the broad‐spectrum phenazine antibiotic d‐alanylgriseoluteic acid.

Metabolomics of extracellular compounds and parasitic enzymes of Beauveria bassiana associated with biological control of whiteflies (Bemisia tabaci)

The Beauveria spp. were isolated from soil and insect cadavers of crop rhizosphere and characterized for parasitic enzyme activity and virulence against whiteflies (Bemisia tabaci). The colony morphology and molecular identification using ITS specific marker were carried out and confirmed entomopathogenic fungi as Beauveria bassiana. The bioefficacy of B. bassiana against whiteflies demonstrated highest corrected mortality and lowest LC50 in isolate B. bassiana JAU2 (SEM morphology) followed by JAU1 on 6th days. Parasitic enzymes chitinase and lipase were determined highest in JAU2 and protease activity examined higher in isolate JAU4 followed by JAU2 isolate on 6th days after inoculation. Comparative extracellular metabolomics carried out from potent (JAU1 and JAU2), moderate (JAU4 and JAU14) and weak (JAU6) B. bassiana isolates in normal suborder dextrose agar with yeast extrect (SDAY) and chitin induced media. Results illustrated that total 105 metabolites identified common for all five B. bassiana isolates differing in virulence. However, the color intensity of the metabolites changes in heat map showing differential concentration of that extracellular compound compared to other isolates. The volcano plot analysis illustrated 58 compounds significanlty diverse between potent JAU1 and JAU2 under two different culture conditions of which 34 compounds recognized up regulated in most potent JAU2 under chitin induced media. Out of 34 metabolites, ten compounds viz., fumaricine, resazurin, N-methyldioctylamine, penaresidun B, tetralin, squamocin B, oligomycin C, pubesenolide, epirbuterol and gentamicin C1a were recognized significantly upregulated in most potent JAU2 and reported for antimicrobial, nematicidal, larvicidalor insecticidal activities. The mass spectra and fragment structure were elucidated under LCMS-QTOF for some novel and unique compounds recognized in most potent B. bassiana JAU2, involved in parasitic activity against whiteflies.