Symbiotic microbiome Staphylococcus aureus from human nasal mucus modulates IL-33-mediated type 2 immune responses in allergic nasal mucosa.

BACKGROUND: The host-microbial commensalism can shape the innate immune responses in respiratory mucosa and nasal microbiome also modulates front-line immune mechanism in the nasal mucosa. Inhaled allergens encounter the host immune system first in the nasal mucosa, and microbial characteristics of nasal mucus directly impact the mechanisms of initial allergic responses in nasal epithelium. However, the roles of the nasal microbiome in allergic nasal mucosa remain uncertain. We sought to determine the distribution of nasal microbiomes in allergic nasal mucosa and elucidate the interplay between nasal microbiome Staphylococcus species and Th2 cytokines in allergic rhinitis (AR) models. RESULTS: Staphylococcus aureus (AR-SA) and S. epidermidis (AR-SE) were isolated from the nasal mucosa of patients with AR. The influence of nasal microbiome Staphylococcus species on allergic nasal mucosa was also tested with in vitro and in vivo AR models. Pyrosequencing data showed that colonization by S. epidermidis and S. aureus was more dominant in nasal mucus of AR subjects. The mRNA and protein levels of IL-33 and TSLP were significantly higher in AR nasal epithelial (ARNE) cells which were cultured from nasal mucosa of AR subjects, and exposure of ARNE cells to AR-SA reduced IL-33 mRNA and secreted protein levels. Particularly, ovalbumin-driven AR mice inoculated with AR-SA by intranasal delivery exhibited significantly reduced IL-33 in their nasal mucosa. In the context of these results, allergic symptoms and Th2 cytokine levels were significantly downregulated after intranasal inoculation of AR-SA in vivo AR mice. CONCLUSION: Colonization by Staphylococcus species was more dominant in allergic nasal mucosa, and nasal commensal S. aureus from subjects with AR mediates anti-allergic effects by modulating IL-33-dependent Th2 inflammation. The results demonstrate the role of host-bacterial commensalism in shaping human allergic inflammation.

Bio-Mediated Synthesis and Characterization of Zinc Phosphate Nanoparticles Using Enterobacter aerogenes Cells for Antibacterial and Anticorrosion Applications.

BACKGROUND: The promising properties of Zinc Phosphate (ZnP) Nanoparticles (NPs) have made them come into prominence as one of the most favorable catalysts in various industries with ever- increasing applications. Among several proposed synthetic methods, biological methods have mostly been desired for their sheer person-environment compatibility in comparison with those of chemical and physical ones. OBJECTIVE: Therefore, the synthesis of ZnP NPs via biological route was developed in this study. METHOD: Herein proposed a facile, applicable procedure for ZnP NPs via biosynthesis route, which included precipitation of Zinc Nitrate (Zn(NO3)2.6H2O) and diammonium hydrogen phosphate ((NH4)2HPO4) in the presence of Enterobacter aerogenes as the synthetic intermediate. Investigation of the anti-corrosion behavior of the synthesized NPs was explored on carbon steel in the hydrochloric acid corrosive environment to provide deeper insight into their unique anti-corrosion properties. Additionally, their antibacterial activities were also examined against Escherichia coli, Staphylococcus aureus and Streptococcus mutans. RESULTS: The results of X-ray Diffraction (XRD), Fourier Transform Infrared (FTIR) spectroscopy, Field Emission Scanning Electron Microscope (FE-SEM) and the Energy Dispersive X-Ray Spectroscopy (EDS) analyses confirmed the successful synthesis of ZnP NPs. Moreover, the examinations of both anti-corrosion and antibacterial properties, revealed that the synthesized NPs could be a promising anti-corrosion/antibacterial agent. CONCLUSION: ZnP NPs with an average size of 30-35 nm were successfully synthesized via the simple, suitable biological method. Results implied that these particles could be used as a non-toxic, environmentally friendly, corrosion-resistant and antibacterial agent instead of toxic and uneco-friendly ones.

Spheroplast-Mediated Carbapenem Tolerance in Gram-Negative Pathogens.

Antibiotic tolerance, the ability to temporarily sustain viability in the presence of bactericidal antibiotics, constitutes an understudied and yet potentially widespread cause of antibiotic treatment failure. We have previously shown that the Gram-negative pathogen Vibrio cholerae can tolerate exposure to the typically bactericidal ß-lactam antibiotics by assuming a spherical morphotype devoid of detectable cell wall material. However, it is unclear how widespread ß-lactam tolerance is. Here, we tested a panel of clinically significant Gram-negative pathogens for their response to the potent, broad-spectrum carbapenem antibiotic meropenem. We show that clinical isolates of Enterobacter cloacae, Klebsiella aerogenes, and Klebsiella pneumoniae, but not Escherichia coli, exhibited moderate to high levels of tolerance of meropenem, both in laboratory growth medium and in human serum. Importantly, tolerance was mediated by cell wall-deficient spheroplasts, which readily recovered wild-type morphology and growth upon removal of antibiotic. Our results suggest that carbapenem tolerance is prevalent in clinically significant bacterial species, and we suggest that this could contribute to treatment failure associated with these organisms.

Hydrogen Production from Energy Poplar Preceded by MEA Pre-Treatment and Enzymatic Hydrolysis.

The need to pre-treat lignocellulosic biomass prior to dark fermentation results primarily from the composition of lignocellulose because lignin hinders the processing of hard wood towards useful products. Hence, in this work a two-step approach for the pre-treatment of energy poplar, including alkaline pre-treatment and enzymatic saccharification followed by fermentation has been studied. Monoethanolamine (MEA) was used as the alkaline catalyst and diatomite immobilized bed enzymes were used during saccharification. The response surface methodology (RSM) method was used to determine the optimal alkaline pre-treatment conditions resulting in the highest values of both total released sugars (TRS) yield and degree of lignin removal. Three variable parameters (temperature, MEA concentration, time) were selected to optimize the alkaline pre-treatment conditions. The research was carried out using the Box-Behnken design. Additionally, the possibility of the re-use of both alkaline as well as enzymatic reagents was investigated. Obtained hydrolysates were subjected to dark fermentation in batch reactors performed by Enterobacter aerogenes ATCC 13048 with a final result of 22.99 mL H2/g energy poplar (0.6 mol H2/mol TRS).

Antimicrobial activity and toxicity of gold nanoparticles: research progress, challenges and prospects.

Gold nanoparticles are emerging materials that exhibit characteristics distinct from those of traditional materials and that have promising potential for application in the fields of chemistry, physics, biology and medicine. During the past decades, numerous studies on the antimicrobial activity and toxicity of gold nanoparticles have been published. With respect to antimicrobial activity, gold nanoparticles conjugated with small molecules, such as antibiotics, drugs, vaccines and antibodies, are more efficient than individual nanoparticles and molecules. Regarding the toxicity effects, results are often unclear and conflicting because of the lack of a standard experimental method; various studies have used different approaches, administration routes and doses, and similar experiments may lead to different conclusions. To provide a systematic overview of and insight in the current knowledge for researchers committed to this filed, we discuss the recent research advances related to the antimicrobial activity and toxicity of gold nanoparticles, both in vitro and in vivo, and identify major issues that require further study. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper discusses the recent research progress on antimicrobial activity and toxicity of gold nanoparticles and provides general insights into the field for researchers committed to this field.

Characterization of Microbial Inactivation Using Plasma-Activated Water and Plasma-Activated Acidified Buffer.

This work investigates the efficacy of plasma-activated water (PAW) and plasma-activated acidified buffer (PAAB) on Enterobacter aerogenes in aqueous system and fruit systems. Reactive oxygen and nitrogen species in PAW have been suggested to provide antimicrobial and acidifying effects, causing the pH of treated water to drop. To isolate the effect of pH in microbial inactivation and to study the interactive effects of pH and reactive species on microbial inactivation, a citrate-phosphate buffer (pH 3.1) and PAAB (citrate-phosphate) were studied. A 1.92 ± 0.70 log CFU/mL reduction in E. aerogenes was observed in PAW, while no reduction was achieved in the buffer, suggesting that the inactivation was due to the reactive species in PAW and not the acidic pH. PAAB achieved a 5.11 ± 0.63 log CFU/mL reduction, suggesting an interactive effect of reactive species and low pH. Electrical conductivity and oxidation-reduction potential measurements suggest potential mechanisms for the greater antimicrobial efficacy of PAAB over PAW. Four surfaces of increasing roughness (glass slides, grape tomatoes, limes, and spiny gourds) were spot inoculated and washed with distilled water, PAW, buffer, and PAAB for 3 min. The smoothest surface (glass) showed the highest reduction (6.32 ± 0.43 log CFU per surface), while the roughest surface (spiny gourd) showed a significantly lower reduction (2.52 ± 0.46 log CFU per surface) when treated with PAAB. For treatment with PAW, no significant differences were observed between glass slides, limes, and spiny gourds. With PAW treatment, significantly lower reduction was observed on spiny gourds (1.70 ± 0.21 log CFU per surface) than on grape tomatoes (4.65 ± 1.34 log CFU per surface). PAW and PAAB both showed potential for their use in fresh produce sanitation.

Boosting mediated electron transfer in bioelectrochemical systems with tailored defined microbial cocultures.

Bioelectrochemical systems (BES) hold great promise for sustainable energy generation via a microbial catalyst from organic matter, for example, from wastewater. To improve current generation in BES, understanding the underlying microbiology of the electrode community is essential. Electron mediator producing microorganism like Pseudomonas aeruginosa play an essential role in efficient electricity generation in BES. These microbes enable even nonelectroactive microorganism like Enterobacter aerogenes to contribute to current production. Together they form a synergistic coculture, where both contribute to community welfare. To use microbial co-operation in BES, the physical and chemical environments provided in the natural habitats of the coculture play a crucial role. Here, we show that synergistic effects in defined cocultures of P. aeruginosa and E. aerogenes can be strongly enhanced toward high current production by adapting process parameters, like pH, temperature, oxygen demand, and substrate requirements. Especially, oxygen was identified as a major factor influencing coculture behavior and optimization of its supply could enhance electric current production over 400%. Furthermore, operating the coculture in fed-batch mode enabled us to obtain very high current densities and to harvest electrical energy for 1 month. In this optimized condition, the coulombic efficiency of the process was boosted to 20%, which is outstanding for mediator-based electron transfer. This study lays the foundation for a rationally designed utilization of cocultures in BES for bioenergy generation from specific wastewaters or for bioprocess sensing and for benefiting from their synergistic effects under controlled bioprocess condition.

Isolation and characterization of antibiotic producing bacterial strains from red soil of Himalayan region of Pakistan.

The emergence of multi drug resistant microbial pathogens has become a global health challenge and set a dire requirement of searching new effective antimicrobials. Soil is an ultimate reservoir of biologically active micro flora, which harbors trillions of microbial strains producing compounds of commercial interest. Hence aim of the present study was an attempt to isolate and identify the antibiotic producing microbial strains from the red soil of Himalayan an unexplored region of Pakistan. In this study from 10 different soil samples only one bacterial strain was isolated capable of antimicrobial activity. Strain was identified by biochemical characteristics and final identification was done by API 20 NE kit which showed 99% homology with P. aeruginosa. Hence the strain was identified as P. aeruginosa S2. Antibacterial and antifungal activity of the P. aeruginosa S2 showed that Staphylococcus aureus was extremely sensitive to it with a zone of inhibition of 42mm. Staphylococcus epidermidis, Enterobacter aerogenes, Aspergillus fumigatus and Candida albicans were also inhibited by the isolated strain. Effect of Glycerol, Copper sulphate (CuSo4), Sodium sulphate (Na2SO4) and Glycerol on antibiotic production was also evaluated by supplementing growth media with these chemicals. Pseudomonas aeruginosa was grown in bulk quantity using solid state fermentation and crude extract was prepared using organic solvents and subjected to silica gel column chromatography for purification of active compound. Purified compound showed antibacterial against human pathogens. The unexplored Kashmir Himalayas are of great significance because of its richness in biodiversity and need to be explored for isolation and characterization of native microbes for biologically active secondary metabolites. This untouched region may be considered as hub of new antimicrobials and may have applications in natural product-based drug discovery.

Enterobacter aerogenes ZDY01 Attenuates Choline-Induced Trimethylamine N-Oxide Levels by Remodeling Gut Microbiota in Mice.

Trimethylamine N-oxide (TMAO), which is transformed from trimethylamine (TMA) through hepatic flavin-containing monooxygenases, can promote atherosclerosis. TMA is produced from dietary carnitine, phosphatidylcholine, and choline via the gut microbes. Previous works have shown that some small molecules, such as allicin, resveratrol, and 3,3-dimethyl-1-butanol, are used to reduce circulating TMAO levels. However, the use of bacteria as an effective therapy to reduce TMAO levels has not been reported. In the present study, 82 isolates were screened from healthy Chinese fecal samples on a basal salt medium supplemented with TMA as the sole carbon source. The isolates belonged to the family Enterobacteriaceae, particularly to genera Klebsiella, Escherichia, Cronobacter, and Enterobacter. Serum TMAO and cecal TMA levels were significantly decreased in choline-fed mice treated with Enterobacter aerogenes ZDY01 compared with those in choline-fed mice treated with phosphate-buffered saline. The proportions of Bacteroidales family S24-7 were significantly increased, whereas the proportions of Helicobacteraceae and Prevotellaceae were significantly decreased through the administration of E. aerogenes ZDY01. Results indicated that the use of probiotics to act directly on the TMA in the gut might be an alternative approach to reduce serum TMAO levels and to prevent the development of atherosclerosis and "fish odor syndrome" through the effect of TMA on the gut microbiota.

Longer Contact Times Increase Cross-Contamination of Enterobacter aerogenes from Surfaces to Food.

Bacterial cross-contamination from surfaces to food can contribute to foodborne disease. The cross-contamination rate of Enterobacter aerogenes on household surfaces was evaluated by using scenarios that differed by surface type, food type, contact time (<1, 5, 30, and 300 s), and inoculum matrix (tryptic soy broth or peptone buffer). The surfaces used were stainless steel, tile, wood, and carpet. The food types were watermelon, bread, bread with butter, and gummy candy. Surfaces (25 cm2) were spot inoculated with 1 ml of inoculum and allowed to dry for 5 h, yielding an approximate concentration of 107 CFU/surface. Foods (with a 16-cm2 contact area) were dropped onto the surfaces from a height of 12.5 cm and left to rest as appropriate. Posttransfer, surfaces and foods were placed in sterile filter bags and homogenized or massaged, diluted, and plated on tryptic soy agar. The transfer rate was quantified as the log percent transfer from the surface to the food. Contact time, food, and surface type all had highly significant effects (P < 0.000001) on the log percent transfer of bacteria. The inoculum matrix (tryptic soy broth or peptone buffer) also had a significant effect on transfer (P = 0.013), and most interaction terms were significant. More bacteria transferred to watermelon (∼0.2 to 97%) than to any other food, while the least bacteria transferred to gummy candy (∼0.1 to 62%). Transfer of bacteria to bread (∼0.02 to 94%) was similar to transfer of bacteria to bread with butter (∼0.02 to 82%), and these transfer rates under a given set of conditions were more variable than with watermelon and gummy candy. IMPORTANCE: The popular notion of the "five-second rule" is that food dropped on the floor and left there for <5 s is "safe" because bacteria need time to transfer. The rule has been explored by a single study in the published literature and on at least two television shows. Results from two academic laboratories have been shared through press releases but remain unpublished. We explored this topic by using four different surfaces (stainless steel, ceramic tile, wood, and carpet), four different foods (watermelon, bread, bread with butter, and gummy candy), four different contact times (<1, 5, 30, and 300 s), and two bacterial preparation methods. Although we found that longer contact times result in more transfer, we also found that other factors, including the nature of the food and the surface, are of equal or greater importance. Some transfer takes place "instantaneously," at times of <1 s, disproving the five-second rule.

Adequate Hand Washing and Glove Use Are Necessary To Reduce Cross-Contamination from Hands with High Bacterial Loads.

Hand washing and glove use are the main methods for reducing bacterial cross-contamination from hands to ready-to-eat food in a food service setting. However, bacterial transfer from hands to gloves is poorly understood, as is the effect of different durations of soap rubbing on bacterial reduction. To assess bacterial transfer from hands to gloves and to compare bacterial transfer rates to food after different soap washing times and glove use, participants' hands were artificially contaminated with Enterobacter aerogenes B199A at ∼9 log CFU. Different soap rubbing times (0, 3, and 20 s), glove use, and tomato dicing activities followed. The bacterial counts in diced tomatoes and on participants' hands and gloves were then analyzed. Different soap rubbing times did not significantly change the amount of bacteria recovered from participants' hands. Dicing tomatoes with bare hands after 20 s of soap rubbing transferred significantly less bacteria (P < 0.01) to tomatoes than did dicing with bare hands after 0 s of soap rubbing. Wearing gloves while dicing greatly reduced the incidence of contaminated tomato samples compared with dicing with bare hands. Increasing soap washing time decreased the incidence of bacteria recovered from outside glove surfaces (P < 0.05). These results highlight that both glove use and adequate hand washing are necessary to reduce bacterial cross-contamination in food service environments.

In vitro antibacterial activities of p-toluenesulfonyl-hydrazinothiazoles and hydrazinoselenazoles against multi-drug resistant Gram-negative phenotypes.

BACKGROUND: Bacterial multidrug resistance (MDR) constitutes a major hurdle in the treatment of infectious diseases worldwide. The present study was designed to evaluate the antibacterial activities of synthetic p-toluenesulfonyl-hydrazinothiazoles against multidrug resistant Gram-negative bacteria. METHODS: The broth microdilution method was used to determine the minimal inhibitory concentrations (MIC). RESULTS: The results demonstrated that the best activities were obtained with hydrazinoselenazoles. p-Chloro-benzyliden-selenosemicarbazide, 4-methyl-2-[(4-chloro-benzyliden)-hydrazinyl]-1,3-selenazole, p-chloro-benzoyl-selenosemicarbazide and 4-chloromethyl-2-[(4-chlorobenziliden)-N-acetyl-hydrazinyl]-1,3-selenazole were more active than the choramphenicol on Klebsiella pneumoniae KP63. Tested alone, the lowest MIC value of 16 mg/L was obtained with p-methoxy-benzyliden-selenosemicarbazide against Enterobacter aerogenes ATCC13048, K. pneumoniae ATCC112296 and KP55. Tested in the presence of an efflux pump inhibitor, phenylalanine arginine ß-naphthylamide (PAßN), the activity of p-chloro-benzyliden-selenosemicarbazide, 4-methyl-2-[(4-chloro-benzyliden)-hydrazinyl]-1,3-selenazole, p-chloro-benzoyl-selenosemicarbazide and p-methoxy-benzyliden-selenosemicarbazide significantly increased with MIC values below 10 mg/L obtained respectively on 43.8 %, 31.3 %, 62.5 % and 100 % of the 16 tested bacterial strains. The lowest MIC value of 0.5 mg/L in the presence of PAßN was recorded with p-chloro-benzoyl-selenosemicarbazide against Escherichia coli ATCC8739 and KP55 as well as p-methoxy-benzyliden-selenosemicarbazide against E. aerogenes KP55. p-Chloro-benzyliden-selenosemicarbazide and p-chloro-benzoyl-selenosemicarbazide contained the same pharmacophore as p-methoxy-benzyliden-selenosemicarbazide. CONCLUSION: This study indicates that p-chloro-benzyliden-selenosemicarbazide, p-chloro-benzoyl-selenosemicarbazide and p-methoxy-benzyliden-selenosemicarbazide could be explored more to develop novel antimicrobial drugs to fight MDR bacterial infections.

Phenolic compounds: Strong inhibitors derived from lignocellulosic hydrolysate for 2,3-butanediol production by Enterobacter aerogenes.

Lignocellulosic biomass are attractive feedstocks for 2,3-butanediol production due to their abundant supply and low price. During the hydrolysis of lignocellulosic biomass, various byproducts are formed and their effects on 2,3-butanediol production were not sufficiently studied compared to ethanol production. Therefore, the effects of compounds derived from lignocellulosic biomass (weak acids, furan derivatives and phenolics) on the cell growth, the 2,3-butanediol production and the enzymes activity involved in 2,3-butanediol production were evaluated using Enterobacter aerogenes ATCC 29007. The phenolic compounds showed the most toxic effects on cell growth, 2,3-butanediol production and enzyme activity, followed by furan derivatives and weak acids. The significant effects were not observed in the presence of acetic acid and formic acid. Also, feasibility of 2,3-butanediol production from lignocellulosic biomass was evaluated using Miscanthus as a feedstock. In the fermentation of Miscanthus hydrolysate, 11.00 g/L of 2,3-butanediol was obtained from 34.62 g/L of reducing sugar. However, 2,3-butanediol was not produced when the concentration of total phenolic compounds in the hydrolysate increased to more than 1.5 g/L. The present study provides useful information to develop strategies for biological production of 2,3-butanediol and to establish biorefinery for biochemicals from lignocellulosic biomass.

Antimicrobial and immunomodulatory efficacy of extracellularly synthesized silver and gold nanoparticles by a novel phosphate solubilizing fungus Bipolaris tetramera.

BACKGROUND: Particulates of nanometers size have occupied a significant area in the field of medicinal and agricultural purposes due to their large surface-to-volume ratio and exceptional physicochemical, electronic and mechanical properties. Myconanotechnology, an interface between mycology and nanotechnology is budding nowadays for nanoparticle-fabrication using fungus or its metabolites. In the present study, we have isolated and characterized a novel phosphate solubilizing fungus B. tetramera KF934408 from rhizospheric soil. This phosphatase releasing fungus was subjected to extracellular synthesis of metal nanoparticles by redox reaction. RESULTS: Silver (AgNPs) and gold nanoparticles (AuNPs) were characterized by dynamic light scattering and transmission electron microscopic analysis. The formulated AgNPs were irregular shaped with a size ranging between 54.78 nm to 73.49 nm whereas AuNPs were spherical or hexagonal, with a size of 58.4 and 261.73 nm, respectively. The nanoparticles were assessed for their antibacterial and antifungal efficacy. The results showed effective antimicrobial activity of AgNPs against Bacillus cereus, Staphylococcus aureus, Enterobacter aeroginosa and Trichoderma sp. at higher concentrations, however, AuNPs possessed only moderate antibacterial efficacy while they found no antifungal activity. Cytotoxicity analysis of nanoparticles on J774 and THP1 α cell lines revealed the dose dependence in case of AgNPs, while AuNPs were non-toxic at both low and high doses. Furthermore, significant elevation of intracellular ROS was observed after 4 h of incubation with both the nanoparticles. The capping of fungal proteins on the particulates might be involved in the activities demonstrated by these inert metal nanoparticles. CONCLUSION: In conclusion, the findings showed that the metal nanoparticles synthesized by fungus B. tetramera could be used as an antimicrobial agents as well as cost effective and nontoxic immunomodulatory delivery vehicle.

Probiotic potential of lactobacillus strains isolated from sorghum-based traditional fermented food.

Sorghum-based traditional fermented food was screened for potential probiotic lactic acid bacteria. The isolates were identified by biochemical, physiological and genetic methods. Species identification was done by 16s rRNA sequence analysis. The functional probiotic potential of the two Lactobacillus species viz., Lactobacillus plantarum (Lact. plantarum) and Lactobacillus pentosus (Lact. pentosus) was assessed by different standard parameters. The strains were tolerant to pH 2 for 1 h and resistant to methicillin, kanamycin, vancomycin and norfloxacin. Two (Lact. plantarum COORG-3 and Lact. pentosus COORG-8) out of eight isolates recorded the cell surface hydrophobicity to be 59.12 and 64.06%, respectively. All the strains showed tolerance to artificial duodenum juice (pH 2) for 3 h, positive for bile salt hydrolase test and negative for haemolytic test. The neutralized cell-free supernatant of the strains Lact. pentosus COORG-4, Lact. plantarum COORG-1, Lact. plantarum COORG-7, Lact. pentosus COORG-8 and Lact. plantarum COORG-3 showed good antibiofilm activity. Lact. pentosus COORG-8 exhibited 74% activity against Pseudomonas aeruginosa-MTCC 7903 and Lact. plantarum COORG-7 showed 68% inhibition of biofilm against Klebsiella pneumonia MTCC 7407. Three (Lact. plantarum COORG-7, Lact. pentosus COORG-5 and Lact. pentosus COORG 8) out of eight isolates exhibited a good antimicrobial activity against Listeria monocytogenes and five isolates (Lact. pentosus COORG 2, Lact. plantarum COORG 1, Lact. plantarum COORG 4, Lact. pentosus COORG 3 and Lact. plantarum COORG 6) are active against Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella pneumonia, Enterococcus faecalis. The study also evaluated the cholesterol lowering property of the Lactobacillus strains using hen egg yolk as the cholesterol source. The cholesterol in hen egg yolk was assimilated by 74.12 and 68.26% by Lact. plantarum COORG 4 and Lact. pentosus COORG 7, respectively. The results of the present study suggest that the Lactobacillus strains isolated and characterized from sorghum-based fermented product may be used as probiotic strains for therapeutic applications.

[MICROBIAL LANDSCAPE OF BILE IN PATIENTS WITH AN ACUTE PURULENT CHOLANGITIS].

Bacteriological analysis was conducted in 136 patients with an acute purulent cholangitis (APCH). The APCH causes were: choledocholithiasis--in 40 (29.9%) patients, coexistence of a common biliary duct stricture and choledocholithiasis--in 39 (28.7%), compression of external biliary ducts by the oedematous pancreatic head in secondary pancreatitis--in 15 (11%), pericholedocheal lymphadenitis--in 3 (2.2%).

Biohydrogen and polyhydroxyalkanoate co-production by Enterobacter aerogenes and Rhodobacter sphaeroides from Calophyllum inophyllum oil cake.

The feasibility of coupled biohydrogen and polyhydroxyalkanoate production by Enterobacter aerogenes and Rhodobacter sphaeroides using Calophyllum inophyllum oil cake was studied under dark and photo fermentation conditions. The utilization of a non-edible acidic oil cake (C. inophyllum), and exploitation of a modified minimal salt media led to reduction in the cost of media. Cost of fermentation is reduced by implementation of alternate dark-photo fermentative periods and through the use of a co-culture consisting of a dark fermentative (E. aerogenes) and a photo fermentative (R. sphaeroides) bacterium. The biohydrogen and polyhydroxyalkanoate produced were 7.95 L H2/L media and 10.73 g/L media, respectively, under alternate dark and photo fermentation and were 3.23 L H2/L media and 5.6g/L media, respectively under complete dark fermentation. The characteristics of the oil cake and alternate dark (16 h) and photo (8h) fermentative conditions were found to be supportive in producing high biohydrogen and polyhydroxyalkanoate (PHA) yield.

Co-fermentation of carbon sources by Enterobacter aerogenes ATCC 29007 to enhance the production of bioethanol.

We investigated the enhancement of bioethanol production in Enterobacter aerogenes ATCC 29007 by co-fermentation of carbon sources such as glycerol, glucose, galactose, sucrose, fructose, xylose, starch, mannitol and citric acid. Biofuel production increases with increasing growth rate of microorganisms; that is why we investigated the optimal growth rate of E. aerogenes ATCC 29007, using mixtures of different carbon sources with glycerol. E. aerogenes ATCC 29007 was incubated in media containing each carbon source and glycerol; growth rate and bioethanol production improved in all cases compared to those in medium containing glycerol alone. The growth rate and bioethanol production were highest with mannitol. Fermentation was carried out at 37 °C for 18 h, pH 7, using 50 mL defined production medium in 100 mL serum bottles at 200 rpm. Bioethanol production under optimized conditions in medium containing 16 g/L mannitol and 20 g/L glycerol increased sixfold (32.10 g/L) than that containing glycerol alone (5.23 g/L) as the carbon source in anaerobic conditions. Similarly, bioethanol production using free cells in continuous co-fermentation also improved (27.28 g/L) when 90.37 % of 16 g/L mannitol and 67.15 % of 20 g/L glycerol were used. Although naturally existing or engineered microorganisms can ferment mixed sugars sequentially, the preferential utilization of glucose to non-glucose sugars often results in lower overall yield and productivity of ethanol. Here, we present new findings in E. aerogenes ATCC 29007 that can be used to improve bioethanol production by simultaneous co-fermentation of glycerol and mannitol.

Volatiles produced by soil-borne endophytic bacteria increase plant pathogen resistance and affect tritrophic interactions.

Volatile organic compounds (VOCs) released by soil microorganisms influence plant growth and pathogen resistance. Yet, very little is known about their influence on herbivores and higher trophic levels. We studied the origin and role of a major bacterial VOC, 2,3-butanediol (2,3-BD), on plant growth, pathogen and herbivore resistance, and the attraction of natural enemies in maize. One of the major contributors to 2,3-BD in the headspace of soil-grown maize seedlings was identified as Enterobacter aerogenes, an endophytic bacterium that colonizes the plants. The production of 2,3-BD by E. aerogenes rendered maize plants more resistant against the Northern corn leaf blight fungus Setosphaeria turcica. On the contrary, E. aerogenes-inoculated plants were less resistant against the caterpillar Spodoptera littoralis. The effect of 2,3-BD on the attraction of the parasitoid Cotesia marginiventris was more variable: 2,3-BD application to the headspace of the plants had no effect on the parasitoids, but application to the soil increased parasitoid attraction. Furthermore, inoculation of seeds with E. aerogenes decreased plant attractiveness, whereas inoculation of soil with a total extract of soil microbes increased parasitoid attraction, suggesting that the effect of 2,3-BD on the parasitoid is indirect and depends on the composition of the microbial community.

Efficacy of knife disinfection techniques in meat processing.

EU Regulation 853/2004 requires that knives used in meat processing be disinfected by submerging them in hot water (+82°C). Alternative procedures are permitted if the efficacy is proved to be equivalent. In the present study, various time-temperature combinations together with pure water, water with lactic acid (2 %), and the use of ultrasound with and without lactic acid (2%) were investigated. Steel plates were covered with fat and protein and then inoculated with a standardized bacterial contamination assembled according to the composition of bacterial contamination found in a previous field trial conducted with regard to the slaughter of pigs. Several combinations with diverse temperatures and time intervals were tested until no microbial load was detectable by using a wet-dry-swab technique that had previously been tested to ensure maximum bacterial recovery. The following were effective in bringing the tested bacterial flora below the detection limit: • 70 °C water bath for 10 s • 60 °C water bath + ultrasound for 5 s • 40 °C water bath + lactic acid (2%) for 10 s • 40 °C water bath + ultrasound + lactic acid (2%) for 5 s In particular, the use of lactic acid permitted a relevant reduction of the temperature, while providing effective sterilization. The use of such non-hazardous food-safe additives is particularly suitable for successful disinfection at lower temperatures.