Exploring the antimicrobial effects of a phenolic-rich extract from jabuticaba depulping waste against enterotoxigenic Escherichia coli.

This study evaluated the effects of a phenolic-rich extract from jabuticaba [Myrciaria jaboticaba (Vell.) Berg] depulping waste (PEJ) on the survival, antibiotic susceptibility, virulence, and cellular functions of various enterotoxigenic Escherichia coli (ETEC) strains. The minimum inhibitory concentration of PEJ against the five tested ETEC strains was 125 mg mL-1. PEJ at 125 and 250 mg mL-1 caused reductions in viable cell counts of ≥ 3 and ≥ 5 log CFU mL-1 in ETEC over 24 h, respectively. PEJ at subinhibitory concentrations (31.25 and 62.5 mg mL-1) reduced the viable cell counts of ETEC when exposed to in vitro gastrointestinal conditions, besides decreasing the biofilm formation, cell surface hydrophobicity, mucin adhesion, and swimming and swarming motility. PEJ (31.25 and 62.5 mg mL-1) increased the susceptibility of the tested ETEC strains to various clinically relevant antibiotics. The exposure to PEJ (62.5 and 125 mg mL-1) impaired the membrane permeability and enzymatic and efflux pump activities in ETEC cells. PEJ effectively reduces survival, increases antibiotic susceptibility, and attenuates virulence in ETEC. These effects could be linked to a PEJ multi-target action disturbing various cellular functions in ETEC cells. PEJ could be a candidate for developing innovative solutions to prevent and treat ETEC infections.

Phenolic-rich extracts from acerola, cashew apple and mango by-products cause diverse inhibitory effects and cell damages on enterotoxigenic Escherichia coli.

This study aimed to evaluate the inhibitory effects of phenolic-rich extracts from acerola (Malpighia emarginata D.C., PEA), cashew apple (Anacardium occidentale L., PEC) and mango (Mangifera indica L., PEM) by-products on distinct enterotoxigenic Escherichia coli (ETEC) strains. The capability of PEA and PEC of impairing various physiological functions of ETEC strains was investigated with multiparametric flow cytometry. Procyanidin B2 , myricetin and p-coumaric acid were the major phenolic compounds in PEA, PEC and PEM, respectively. PEA and PEC had lower minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) (MIC: 31·25 mg ml-1 ; MBC: 62·5 mg ml-1 ) on ETEC strains than PEM (MIC and MIC: >1000 mg ml-1 ). PEA and PEC (15·6, 31·2, 62·5 mg ml-1 ) caused viable count reductions (P < 0·05) on ETEC strains after 24 h of exposure, notably the ≥3 log reductions caused by 62·5 mg ml-1 . The 24 h exposure of ETEC strains to PEA and PEC (31·2, 62·5 mg ml-1 ) led to high sizes of cell subpopulations with concomitant impairments in cell membrane polarization and permeability, as well as in enzymatic, respiratory and efflux activities. PEA and PEC are effective in inhibiting ETEC through a multi-target action mode with disturbance in different physiological functions.

A review of the current evidence of fruit phenolic compounds as potential antimicrobials against pathogenic bacteria.

Fruits are among the main natural sources of phenolic compounds (PC). These compounds exert important antioxidant properties primarily associated with the presence of hydroxyl groups in their molecular structure. Additionally, the antibacterial effects of fruit phenolic-rich extracts or individual PC commonly found in fruits have been an emerging research focus in recent years. This review discusses by first time the available literature regarding the inhibitory effects of fruit PC on pathogenic bacteria, including not only their direct effects on bacterial growth and survival, but also their effects on virulence factors and antibiotic resistance, as well as the possible mechanism underlying these inhibitory properties. The results of the retrieved studies show overall that the antibacterial effects of fruit PC vary with the target bacteria, type of PC and length of exposure to these compounds. The type of solvent and procedures used for extraction and fruit cultivar also seem to influence the antibacterial effects of phenolic-rich fruit extracts. Fruit PC have shown wide-spectrum antibacterial properties besides being effective antibiotic resistance modifying agents in pathogenic bacteria and these effects have shown to be associated with interruption of efflux pump expression/function. Furthermore, fruit PC can cause down regulation of a variety of genes associated with virulence features in pathogenic bacteria. Results of available studies indicate the depolarization and alteration of membrane fluidity as mechanisms underlying the inhibition of pathogenic bacteria by fruit PC. These data reveal fruit PC have potential antimicrobial properties, which should be rationally exploited in solutions to control pathogenic bacteria.

Enterotoxigenic Escherichia coli targeting intestinal epithelial tight junctions: An effective way to alter the barrier integrity.

Enterotoxigenic Escherichia coli are responsible for causing secretory diarrhea in animal(s), including human(s). This group of microorganisms is classified on the basis of production of toxins acting on the intestinal epithelium of the small intestine. Various enterotoxins, heat-labile and heat-resistant, are produced by distinct strains of ETEC. Although the mechanisms of action of ETEC enterotoxins were shown to involve diverse ion channels recent data suggest that these molecules could also be involved in disruption of the permeability barrier of the intestinal epithelium. More precisely, the tight junctions directly responsible for the selective permeability of the intestinal tissue could be affected. Studies indicating a change in TJ following exposure of cell monolayers or animal models either to pure enterotoxins or to ETEC strains producing one or more of these toxic molecules will be discussed.

[Biochemical recurrence after curative treatment for localized prostate cancer: Performance of choline PET/CT in the assessment of local recurrence]. / Récidive biochimique après traitement curatif d'un adénocarcinome prostatique localisé: intérêt de la TEP à la choline dans l'évaluation de la récidive locale.

OBJECTIVE: To establish 18 fluorocholine-positron emission tomography/computed tomography (F-PET/CT) performances for the detection of local recurrence in a population of patients with biochemical failure after primary curative treatment for localized prostate carcinoma. MATERIAL AND METHOD: From February 2011 to February 2014, 55 patients underwent a F-PET/CT for biochemical relapse after primary radical therapy for prostate cancer localized or locally advanced. Primary therapies for prostate cancer were 19 radical prostatectomy, 18 radiotherapy, 13 radiotherapy with hormonal treatment, 3 brachytherapy. The median age was 65 years (50-79). The initial staging was 17 T1, 23 T2 and 15 T3, 52 were N0 and N1 3. The median PSA was 12 (3-127). The Gleason score was less than 7, equal to 7 and greater than 7 at 21, 25 and 9 patients respectively. The average time to recurrence was 69.5 months (8-147) with a median PSA of 2.9 ng/mL (0.48-41). RESULTS: In 42 cases, F-PET/CT showed uptake, suggesting a recurrence, metastatic (6), nodal (26) or local isolated (10). The focal uptake in PET commissioned in 5 cases prostate biopsy, confirming the histological recurrence of prostate cancer in 4 cases. Among the 10 patients with isolated local recurrence, 8 underwent salvage radiotherapy. Of the 13 cases where the (F-PET/CT) showed no recurrence, 7 multiparametric MRI were performed. The MRI showed a local recurrence in 3 patients, the diagnoses were confirmed with prostate biopsy for two of them. CONCLUSION: In our study, for the patients with biochemical relapse of prostate adenocarcinoma localized or locally advanced, (F-PET/CT) was able to detect local recurrence isolated in nearly half the cases but did not show sufficient sensitivity to exclude recurrence local if negative. It does not replace MRI or additional prostate biopsy.

Escherichia coli heat-stable toxin b impairs intestinal epithelial barrier function by altering tight junction proteins.

Escherichia coli heat-stable toxin b (STb) causes diarrhea in animals. STb binds to sulfatide, its receptor, and is then internalized. In the cytoplasm, through a cascade of events, STb triggers the opening of ion channels, allowing ion secretion and water loss and leading to diarrhea. Tight junctions (TJs) are well known for controlling paracellular traffic of ions and water by forming a physical intercellular barrier in epithelial cells, and some bacterial toxins are known to affect adversely TJs. The present study aimed at determining the effect of STb on TJs. T84 cells were treated for 24 h with purified STb and a nontoxic STb mutant (D30V). Transepithelial resistance (TER), paracellular flux marker, and confocal microscopy were used to analyze the effect of STb on TJs. Purified STb caused a significant reduction of TER parallel to an increase in paracellular permeability compared to the results seen in untreated cells or mutant D30V. The increased paracellular permeability was associated with a marked alteration of F-actin stress fibers. F-actin filament dissolution and condensation were accompanied by redistribution and/or fragmentation of ZO-1, claudin-1, and occludin. These changes were also observed following treatment of T84 cells with an 8-amino-acid peptide found in the STb sequence corresponding to a consensus sequence of Vibrio cholerae Zot toxin. These effects were not observed with a scrambled peptide or mutant D30V. Our findings indicate that STb induces epithelial barrier dysfunction through changes in TJ proteins that could contribute to diarrhea.

Membrane filtration of the liquid fraction from a solid-liquid separator for swine manure using a cationic polymer as flocculating agent.

The liquid fraction from a solid-liquid separator for swine manure, which used a cationic polymer to promote particle flocculation, was processed by one nanofiltration and two reverse osmosis spiral-wound membranes. Eight different liquid fraction batches (750 to 1750 L) were concentrated at volumetric concentration ratios (VCRs, initial to final volumes) ranging from 2.3 to 4.2. Membrane fouling intensity was highly variable, as water flux recovery after concentration cycles ranged from 13% to 88%. The most severe fouling was caused by a liquid fraction that had relatively low suspended solids (SS) (774 mg/L) and was concentrated at a low VCR of 2.6. Raw manure collected the same day also contained low SS, suggesting that fewer sites were available for polymer adsorption and thus more polymer remained in the liquid. However, because of the high opacity of the samples, residual polymer could not be detected in any feed or concentrate samples. Fouling was not totally irreversible as over 97% of membrane flux could be recovered by cleaning with acidic and alkaline solutions. Further tests with spiked liquid fractions indicated that fouling due to residual polymer in solution started to occur at a polymer concentration of 3 and 11 mg/L in initial and concentrated effluents, respectively. If a cationic polymer is used to pretreat manure, the amount of added polymer would have to be closely related to SS content as opposed to manure volume, in order to leave very little residual polymer in solution.

The whole Shebang: the gastrointestinal tract, Escherichia coli enterotoxins and secretion.

This review focuses on diarrhea caused by toxins released by enterotoxigenic Escherichia coli. These bacteria are known to produce toxins that have adverse effects on the intestinal tissue in Man and animals. E. coli is contracted through the ingestion of water or food contaminated by this bacterium. Generally, E. coli colonizes the intestinal mucosa where it multiplies and causes damage to the target cells or interferes with the homeostasis that prevails in the gastrointestinal tract. Enteropathogens such as E. coli are only able to exhibit their effects after colonization of the intestinal mucosa from where they release their toxins. These bacteria mainly affect chloride ions secretion through second messenger pathways resulting in secretory diarrhea. In this review, the association of bacteria with the gastrointestinal tract as pathogens and the resulting effects on the various systems of the intestine, including the nervous system and mediators leading to secretion and diarrhea are examined.

Escherichia coli STb toxin induces apoptosis in intestinal epithelial cell lines.

A previous study conducted in our laboratory demonstrated that cells having internalized Escherichia coli STb toxin display apoptotic-like morphology. We therefore investigated if STb could induce programmed cell death in both a human and an animal intestinal epithelial cell lines. HRT-18 (Human Colon Tumor) and IEC-18 (Rat Ileum Epithelial Cells) cell lines were used. As STb is frequently tested in a rat model, the IEC-18 cell line was most relevant to our work. The cell lines were treated with various amounts of purified STb (nanomole range) for a period of 24 h after which cells were harvested and examined for apoptotic characteristics. Caspase-9, the initiator of mitochondrion-mediated apoptosis, and caspase-3, an effector of caspase-9, were both activated following STb intoxication of HRT-18 and IEC-18 cells whereas caspase-8, the initiator caspase of the extrinsic pathway, was not activated. For both cell lines, agarose gel electrophoresis of the cell DNA content reveals laddering of DNA, resulting from DNA fragmentation, a characteristic of apoptosis. Hoechst 33342-stained DNA of STb-treated cell lines, observed using fluorescence microscopy, revealed condensation and fragmentation of the nuclei. Apoptotic indexes calculated from fragmented nuclei of Hoechst 33342-stained DNA for HRT-18 and IEC-18 cells showed an STb dose-dependent response. Overall, these data indicate that STb toxin induces a mitochondrion-mediated caspase-dependent apoptotic pathway.

Nitrogen potential recovery and concentration of ammonia from swine manure using electrodialysis coupled with air stripping.

The practice of intensive animal production in certain areas has resulted in excessive manure production for the available regional land base. Consequently, there is a need to develop treatment technologies to recover the valuable nutrients that manure contains so that the resulting product can be transported and used as fertilizer on agricultural land. The project presented here used electrodialysis in a dilution/concentration configuration to transfer the manure ammonia in the diluate solution by electromigration to an adjacent solution separated by an ion-exchange membrane under the driving force of an electrical potential. Then, air stripping from the electrodialysis-obtained concentrate solution without pH modification was used to isolate the ammonia in an acidic solution. An optimal process operating voltage of 17.5 V was first determined on the basis of current efficiency and total energy consumption. During the process, the swine manure pH varied from 8.5 to 8.2, values favourable for NH(4)(+) electromigration. Total ammonia nitrogen reached 21,352 mg/L in the concentrate solution, representing approximately seven times the concentration in the swine manure. Further increases in concentration were limited by water transfer from the diluate solution due to electroosmosis and osmosis. Applying vacuum to the concentrate reservoir was found to be more efficient than direct concentrate solution aeration for NH(3) recuperation in the acid trap, given that the ammonia recuperated under vacuum represented 14.5% of the theoretical value of the NH(3) present in the concentrate solution as compared to 6.2% for aeration. However, an excessively low concentrate solution pH (8.6-8.3) limited NH(3)volatilization toward the acid trap. These results suggest that the concentrate solution pH needs to be raised to promote the volatile NH(3) form of total ammonia nitrogen.

Pig vaccination strategies based on enterotoxigenic Escherichia coli toxins.

Enterotoxigenic Escherichia coli (ETEC) are responsible for diarrhea in humans as well as in farm animals. ETEC infections in newborn, suckling, and especially in post-weaning piglets are associated with reduced growth rate, morbidity, and mortality. ETEC express virulence factors as adhesin and enterotoxins that play a central role in the pathogenic process. Adhesins associated with pigs are of diverse type being either fimbrial or non-fimbrial. Enterotoxins belong to two groups: heat-labile (LT) and heat-stable (ST). Heterogeneity of ETEC strains encompass expression of various fimbriae (F4, F5, F6, F18, and F41) and enterotoxins (LT, STa, STb, and EAST1). In the late years, attempts to immunize animals against neonatal and post-weaning diarrhea were focused on the development of anti-adhesin strategies as this is the initial step of ETEC pathogenesis. Although those vaccines demonstrated some protection against ETEC infections, as enterotoxins are pivotal to the virulence of ETEC, a new generation of vaccinal molecules, which include adhesin and one or more enterotoxins, were recently tested. Some of these newly developed chimeric fusion proteins are intended to control as well human diarrhea as enterotoxins are more or less common with the ones found in pigs. As these could not be tested in the natural host (human), either a mouse or pig model was substituted to evaluate the protection efficacy. For the advancement of pig vaccine, mice were sometimes used for preliminary testing. This review summarizes advances in the anti-enterotoxin immunization strategies considered in the last 10 years.

STb and AIDA-I: the missing link?

Escherichia coli enterotoxigenic strains produce one or more toxins which action result in production of diarrhea in animals including Man. One of these toxins, STb, has been mainly associated with colibacillosis in swine. Although highly prevalent in pigs with diarrhea, a relation between STb and disease was arduous to establish. With the recent recognition of a new adhesin, originally found in human E. coli isolates, named AIDA (adhesin involved in diffuse adherence) and its association with new E. coli pathotypes to which STb is linked, new light was shed on STb toxic potency. In this review, the association of STb and AIDA is examined according to the recent knowledge gained with newly described E. coli pathotypes.

Fruit extracts to control pathogenic Escherichia coli: A sweet solution.

Escherichia coli is a major cause of diarrhea and is as well responsible for extraintestinal infections in humans and animals. Many pathotypes have been defined for this ubiquitous microorganism on the basis of the virulence attributes. For the last 70 years, antibiotics have been used to control infections caused by E. coli. However, with the resistance observed with many strains these drugs are less recommended. Plant extracts, in particular fruit, represent a source of bioactive compounds that could be beneficial in the control of infectious diseases caused by E. coli. These could have bacteriostatic or bactericidal potential or could be used as synergic agents to amplify the activity of antibiotics for which the germs present some level of resistance. Certain studies also revealed that fruit extracts could act directly on virulence characters to attenuate the pathogenic capacity of microorganisms. This review intent to expose the scant but rapidly growing information available that shows that fruit, used as crude extracts or purified molecules, should be considered to manage diverse types of infections caused by E. coli.

Energy density of anchovy Engraulis encrasicolus in the Bay of Biscay.

The energy density (E(D)) of anchovy Engraulis encrasicolus in the Bay of Biscay was determined by direct calorimetry and its evolution with size, age and season was investigated. The water content and energy density varied seasonally following opposite trends. The E(D) g(-1) of wet mass (M(W)) was highest at the end of the feeding season (autumn: c. 8 kJ g(-1)M(W)) and lowest in late winter (c. 6 kJ g(-1)M(W)). In winter, the fish lost mass, which was partially replaced by water, and the energy density decreased. These variations in water content and organic matter content may have implications on the buoyancy of the fish. The water content was the major driver of the energy density variations for a M(W) basis. A significant linear relationship was established between E(D) g(-1) (y) and the per cent dry mass (M(D); x): y =-4.937 + 0.411x. In the light of the current literature, this relationship seemed to be not only species specific but also ecosystem specific. Calibration and validation of fish bioenergetics models require energy content measurements on fish samples collected at sea. The present study provides a first reference for the energetics of E. encrasicolus in the Bay of Biscay.

EAST1 toxin: An enigmatic molecule associated with sporadic episodes of diarrhea in humans and animals.

EAST1 is produced by a subset of enteroaggregative Escherichia coli strains. This toxin is a 38-amino acid peptide of 4100 Da. It shares 50% homology with the enterotoxic domain of STa and interacts with the same receptor. The mechanism of action of EAST1is proposed to be identical to that of STa eliciting a cGMP increase. EAST1 is associated with diarrheal disease in Man and various animal species including cattle and swine. Nevertheless, as EAST1-positive strains as well as culture supernatants did not provoke unequivocally diarrhea either in animal models or in human volunteers, the role of this toxin in disease is today still debated. This review intent is to examine the role of EAST1 toxin in diarrheal illnesses.

Helicobacter pylori interactions with host serum and extracellular matrix proteins: potential role in the infectious process.

Helicobacter pylori, a gram-negative spiral-shaped bacterium, specifically colonizes the stomachs of humans. Once established in this harsh ecological niche, it remains there virtually for the entire life of the host. To date, numerous virulence factors responsible for gastric colonization, survival, and tissue damage have been described for this bacterium. Nevertheless, a critical feature of H. pylori is its ability to establish a long-lasting infection. In fact, although good humoral (against many bacterial proteins) and cellular responses are observed, most infected persons are unable to eradicate the infection. A large body of evidence has shown that the interaction between H. pylori and the host is very complex. In addition to the effect of virulence factors on colonization and persistence, binding of specialized bacterial proteins, known as receptins, to certain host molecules (ligands) could explain the success of H. pylori as a chronically persisting pathogen. Some of the reported interactions are of high affinity, as revealed by their calculated dissociation constant. This review examines the binding of host proteins (serum and extracellular matrix proteins) to H. pylori and considers the significance of these interactions in the infectious process. A more thorough understanding of the kinetics of these receptin interactions could provide a new approach to preventing deeper tissue invasion in H. pylori infections and could represent an alternative to antibiotic treatment.

Modulation of hexa-acyl pyrophosphate lipid A population under Escherichia coli phosphate (Pho) regulon activation.

Environmental phosphate is an important signal for microorganism gene regulation, and it has recently been shown to trigger some key bacterial virulence mechanisms. In many bacteria, the Pho regulon is the major circuit involved in adaptation to phosphate limitation. The Pho regulon is controlled jointly by the two-component regulatory system PhoR/PhoB and by the phosphate-specific transport (Pst) system, which both belong to the Pho regulon. We showed that a pst mutation results in virulence attenuation in extraintestinal pathogenic Escherichia coli (ExPEC) strains. Our results indicate that the bacterial cell surface of the pst mutants is altered. In this study, we show that pst mutants of ExPEC strains display an increased sensitivity to different cationic antimicrobial peptides and vancomycin. Remarkably, the hexa-acylated 1-pyrophosphate form of lipid A is significantly less abundant in pst mutants. Among differentially expressed genes in the pst mutant, lpxT coding for an enzyme that transfers a phosphoryl group to lipid A, forming the 1-diphosphate species, was found to be downregulated. Our results strongly suggest that the Pho regulon is involved in lipid A modifications, which could contribute to bacterial surface perturbations. Since the Pho regulon and the Pst system are conserved in many bacteria, such a lipid A modification mechanism could be widely distributed among gram-negative bacterial species.

Escherichia coli STb toxin and colibacillosis: knowing is half the battle.

Expression of both adherence and enterotoxin expression are required for enterotoxigenic Escherichia coli (ETEC) strains to cause colibacillosis. ETEC strains are responsible for diarrhea in humans and animals by production of various enterotoxins. For many years, the role of the heat-stable E. coli enterotoxin STb as a diarrhea-causing toxin in animals, and in particular in swine, has been controversial. In fact, although the presence of STb-positive E. coli strains and diarrhea in animals is frequently observed, the difficulty of reproducing the pathology in an animal model was interpreted as a lack of toxicity. Recently, new light was shed on the activity of STb in intestinal ligated loops and in pigs orally inoculated with STb-positive E. coli strains. This minireview revisits the effects of STb on the intestinal epithelium and enlightens the significance of STb in swine colibacillosis. The interaction of STb toxin with other E. coli enterotoxins and dual ETEC/enteropathogenic E. coli or ETEC/attaching effacing E. coli infections are also discussed.

Escherichia coli STb toxin binding to sulfatide and its inhibition by carragenan.

Escherichia coli heat-STb is an important cause of diarrhea in piglets. STb was shown to interact specifically with sulfatide (3'-sulfogalactosyl-ceramide) present on the surface of epithelial cells of piglet jejunum. Basic data are lacking on STb binding to sulfatide in solution and more precisely on the possible inhibition of this interaction. Using surface plasmon resonance technology, we compare binding of STb to sulfatide and other glycoshingolipids previously shown, with a multiplate-binding assay, to also interact to various degrees with the enterotoxin. In addition, inhibition of STb-sulfatide binding was studied using free galactose, galactose-sulfate residues and a polymer of sulfated galactans known as carragenan. We determined a dissociation constant of 2.4+/-0.61 nM for the STb-sulfatide interaction. These data indicated that STb was binding to sulfatide with greater affinity than previously determined using radiolabeled toxin. Much lower affinities were observed for lactoceramide and glucoceramide. The binding of STb to sulfatide was clearly inhibited by lambda-carragenan but not by galactose, 4-SO(4)-galactose or 6-SO(4)-galactose. Inhibition of STb binding to its receptor was achieved using lambda-carragenan at picomolar concentrations. Then, using IPEC-J2 cells in culture and flow cytometry, we showed that lambda-carragenan was able to inhibit the permeabilization process associated with STb.