Campylobacter devanensis sp. nov., Campylobacter porcelli sp. nov., and Campylobacter vicugnae sp. nov., three novel Campylobacter lanienae-like species recovered from swine, small ruminants, and camelids.

In a previous study characterizing Campylobacter strains deficient in selenium metabolism, 50 strains were found to be similar to, but distinct from, the selenonegative species Campylobacter lanienae. Initial characterization based on multilocus sequence typing and the phylogeny of a set of 20 core genes determined that these strains form three putative taxa within the selenonegative cluster. A polyphasic study was undertaken here to further clarify their taxonomic position within the genus. The 50 selenonegative strains underwent phylogenetic analyses based on the sequences of the 16S rRNA gene and an expanded set of 330 core genes. Standard phenotypic testing was also performed. All strains were microaerobic and anaerobic, Gram-negative, spiral or curved cells with some displaying coccoid morphologies. Strains were motile, oxidase, catalase, and alkaline phosphatase positive, urease negative, and reduced nitrate. Strains within each clade had unique phenotypic profiles that distinguished them from other members of the genus. Core genome phylogeny clearly placed the 50 strains into three clades. Pairwise average nucleotide identity and digital DNA-DNA hybridization values were all below the recommended cut-offs for species delineation with respect to C. lanienae and other related Campylobacter species. The data presented here clearly show that these strains represent three novel species within the genus, for which the names Campylobacter devanensis sp. nov. (type strain RM3662T=LMG 33097T=NCTC 15074T), Campylobacter porcelli sp. nov. (type strain RM6137T=LMG 33098T=CCUG 77054T=NCTC 15075T) and Campylobacter vicugnae sp. nov. (type strain RM12175T=LMG 33099T=CCUG 77055T=NCTC 15076T) are proposed.

Sequencing and functional validation of the JGI Brachypodium distachyon T-DNA collection.

Due to a large and growing collection of genomic and experimental resources, Brachypodium distachyon has emerged as a powerful experimental model for the grasses. To add to these resources we sequenced 21 165 T-DNA lines, 15 569 of which were produced in this study. This increased the number of unique insertion sites in the T-DNA collection by 21 078, bringing the overall total to 26 112. Thirty-seven per cent (9754) of these insertion sites are within genes (including untranslated regions and introns) and 28% (7217) are within 500 bp of a gene. Approximately 31% of the genes in the v.2.1 annotation have been tagged in this population. To demonstrate the utility of this collection, we phenotypically characterized six T-DNA lines with insertions in genes previously shown in other systems to be involved in cellulose biosynthesis, hemicellulose biosynthesis, secondary cell wall development, DNA damage repair, wax biosynthesis and chloroplast synthesis. In all cases, the phenotypes observed supported previous studies, demonstrating the utility of this collection for plant functional genomics. The Brachypodium T-DNA collection can be accessed at http://jgi.doe.gov/our-science/science-programs/plant-genomics/brachypodium/brachypodium-t-dna-collection/.

Antimicrobial Poly(lactic acid)-Based Nanofibres Developed by Solution Blow Spinning.

The present study reports on the development of hybrid poly(lactic acid) (PLA) fibres loaded with highly crystalline bacterial cellulose nanowhiskers (BCNW) by the novel solution blow spinning method. Furthermore, fibres with antimicrobial properties were generated by incorporating carvacrol and THC as antimicrobial agents and the biocide effect against Listeria monocytogenes was studied. Initially, PLA blow spun fibres containing BCNW were optimized in terms of morphology and thermal properties. The addition of BCNW was seen to significantly increase the viscosity and surface tension of solutions, restricting the capacity to form fibres for concentrations greater than 30 wt.-% BCNW. 15 wt.-% BCNW was selected as the optimum nanofiller loading as it led to the most uniform fibres morphology, with BCNW homogeneously distributed along the fibres' axis. Subsequently, carvacrol and THC were incorporated into the fibres to confer them with antimicrobial properties, although the hydrophobic PLA matrix did not provide an efficient release of the antimicrobials. Thus, hydrophilic substances were added in order to trigger the antimicrobials release through water sorption mechanisms. The addition of the BCNW filler was not seen to significantly increase the antimicrobial capacity of the fibres by itself and, hence, gelatin was added to help promoting further the hydrophylicity and biocide performance of the fibres. Nevertheless, for the more hydrophilic THC, the biocide capacity of the fibres with gelatin was accentuated further by the presence of the BCNW.

Potential of NIR spectroscopy for predicting cellulose nanofibril quality in commercial bleached Kraft pulp of Eucalyptus.

Multivariate models were developed to classify cellulose nanofibril (CNF) fibrillation by a quality index from near infrared (NIR) spectra. Commercial pulps of Eucalyptus spp. were used to produce cellulose nanofibrils by means of a fibrillator mill. After each of the five passes through the mill, samples were collected and analyzed for energy consumption and fiber classification. As a standard, pulps were oxidized with TEMPO reagent followed by a single pass through the mill to compare the resulting quality of CNFs produced by each method. NIR spectra of CNFs were associated with quality indices determined by conventional laboratory analyses that included morphology, turbidity, mechanical properties, X-ray diffraction and quality index measurements. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were applied to the spectral and experimental data. Fibrillator milling to obtain CNFs was efficient and resulted in gel formation following the third pass through the mill. NIR spectroscopy combined with PLS-DA was used successfully to create a model to classify quality of CNFs with 96 % certainty in 3 wt% solutions. These findings suggest that NIR spectroscopy holds promise for estimating CNF quality in suspension, particularly in real-time industrial applications where reliable estimates are crucial.

Bio-based thin films of cellulose nanofibrils and magnetite for potential application in green electronics.

The objective of this work was to prepare bio-based thin films and evaluate the additions of magnetite and glycerol on the physico-chemical (flexibility, wettability and barrier properties) and dielectric properties of cellulose/chitosan-based films. The films were prepared by solution casting and presented a suitable dispersion of the constituents observed by SEM and FTIR. The films were thermally stable up to 150 °C and had a higher flexibility, wettability and lower barrier properties upon addition of glycerol. The calculated dielectric constant (εr) for the composite films was based on measurements of capacitance, at 100 and 1000 Hz, with the additions of magnetite and glycerol more than doubling the εr increasing the charge storage capacity. The bio-based thin films have potential to be used as insulators in capacitors on the production of green electronics thus, reducing toxic and nonrenewable e-waste generation.

Massaranduba Sawdust: A Potential Source of Charcoal and Activated Carbon.

This paper provides proof of concept that activated carbon (AC) may be readily produced using limited conversion methods and resources from sawdust of massaranduba (Manilkara huberi) wood, thereby obtaining value-added products. Sawdust was sieved and heat-treated in an oxygen-free muffle furnace at 500 °C to produce charcoal. The charcoal was activated in a tubular electric furnace at 850 °C while being purged with CO2 gas. Microstructural, thermal and physical properties of the three components: sawdust, charcoal and AC were compared by means of field emission scanning electron microscopy (FESEM), X-ray diffractometry (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), density and water adsorption/desorption measurements. The resulting AC had a large surface area as measured by Brunauer-Emmett-Teller (BET) comparable to other such values found in the literature. The large surface area was due to pore development at the microstructural level as shown by FESEM. XRD illustrated that sawdust had a semi-crystalline structure whereas charcoal and AC evidenced mostly amorphous structures. TGA and DSC showed that AC had high reactivity to moisture compared to sawdust and charcoal.

Renewable hybrid nanocatalyst from magnetite and cellulose for treatment of textile effluents.

A hybrid catalyst was prepared using cellulose nanofibrils and magnetite to degrade organic compounds. Cellulose nanofibrils were isolated by mechanical defibrillation producing a suspension used as a matrix for magnetite particles. The solution of nanofibrils and magnetite was dried and milled resulting in a catalyst with a 1:1 ratio of cellulose and magnetite that was chemically and physically characterized using light, scanning electron and transmission electron microscopies, specific surface area analysis, vibrating sample magnetometry, thermogravimetric analysis, Fourier transform infrared spectroscopy, X-ray diffraction, catalytic potential and degradation kinetics. Results showed good dispersion of the active phase, magnetite, in the mat of cellulosic nanofibrils. Leaching and re-use tests showed that catalytic activity was not lost over several cycles. The hybrid material produced was tested for degradation of methylene blue dye in Fenton-like reactions resulting in a potential catalyst for use in degradation of organic compounds.

Preparation of Zein Fibers Using Solution Blow Spinning Method.

Zein fibers were successfully fabricated via solution blow spinning (SBS) using acetic acid as solvent. Surface tension, viscosity and modulus of zein solutions were respectively determined by force tensiometer and rheometer. Increases of these properties were observed with an increase of concentration from 20% to 35% (w/w). The fabrication conditions of zein fibers were initially investigated as a function of zein concentration (25% to 35% w/w), feed rate (0.04 to 0.1 mL/min) and air pressure (0.28 to 0.62 MPa). The average fiber diameter (AFD) ranged from 174 to 9595 nm based on scanning electron microscopy (SEM). A Box-Behnken experimental design (BBD) was further performed to identify and quantify the significance of above parameters. The statistical analysis showed that the linear coefficient of concentration, the quadratic term of concentration as well as the interaction between concentration and air pressure were demonstrated statistically significant. Optimal conditions, with an AFD of 138 nm, could be obtained in the SBS of zein fibers by combining a concentration of 23% (w/w), a feed rate of 0.04 mL/min and an air pressure of 0.38 MPa. The moisture sorption capacity of fibers increased slightly as AFD decreased from ∼550 to ∼200 nm, with an increase of BET surface area from 116.5 to 140.0 m2 /g.

Physical and antibacterial properties of açaí edible films formulated with thyme essential oil and apple skin polyphenols.

UNLABELLED: Thyme essential oil (TEO) and apple skin polyphenols (ASP) are natural compounds considered as generally recognized as safe by FDA, with biological effects against bacteria and fungi. This work aimed to evaluate physical and antimicrobial properties of açaí edible films formulated with TEO and ASP at 3% and 6% (w/w) individually or combined at 3% (w/w) each. Physical properties studied include mechanical resistance, water vapor permeability (WVP), color, and thermal resistance. Antimicrobial activity against Listeria monocytogenes was determined using the overlay diffusion test. Addition of ASP resulted in improved mechanical properties. TEO at 6% (w/w) resulted in increased elongation. ASP films had significant higher WVP than control film. ASP films were lighter and had more red color than other films. Incorporation of ASP resulted in improved film thermal stability, whereas TEO caused rapid thermal decomposition. Presence of clusters was observed on the surface of films. Addition of ASP resulted in a smoother surface, whereas addition of TEO led to the formation of crater-like pits on the film surface. Açaí edible film incorporated with 6% (w/w) TEO presented the highest antimicrobial activity. However, both antimicrobials are necessary in the açaí films in order to obtain edible films with suitable physical-mechanical properties. The results of the present study showed that TEO and ASP can be used to prepare açaí edible films with adequate physical-mechanical properties and antimicrobial activity for food applications by direct contact. PRACTICAL APPLICATION: Developed açaí edible films presented antimicrobial activity against L. monocytogenes and good physical-mechanical properties, showing the potential use of açaí edible films in food preservation.

Solid lipid particles in lipid films to control the diffusive release of 2-heptanone.

BACKGROUND: Controlled-release formulations of bioactive agents are of increasing interest for effective pest control. Volatile 2-heptanone is a bioactive agent that has shown potential as a pesticide. The aim of this study was to investigate the kinetics of release of 2-heptanone incorporated into lipid films or composite solid lipid particle (SLP) films. RESULTS: Effective 2-heptanone diffusivity was estimated to be between 0.1 and 2.5 mm(2) day(-1) during the first week and between 0.05 and 0.1 mm(2) day(-1) during the next 5 weeks. The films that showed better retention of 2-heptanone were the paraffin lipid films. Inclusion of SLPs into paraffin films increased the release rate of 2-heptanone, mainly owing to a decrease in the film firmness as the composite SLP film became less crystalline and more brittle. In contrast, SLPs decreased the kinetics of 2-heptanone release in Acetem films owing to an increase in the film firmness. CONCLUSIONS: The results indicated that the use of SLPs as a method for controlled release can improve the delivery of the natural pesticide 2-heptanone if the SLPs have good compatibility with the matrix, leading to an increase in firmness of the films without increasing their porosity. Published 2012. This article is a U.S. Government work and is in the public domain in the USA.

Use of microscopy to assess bran removal patterns in milled rice.

During rice milling, the bran and germ are successively removed from the caryopsis (kernel). Because bran and germ contain large quantities of lipid, the amount of lipid remaining on the kernel surface may be used as a method for the assessment of milling quality. Bulk samples of rice pureline varieties and an experimental hybrid were milled for 0, 10, 20, 30, and 40 s. Scanning electron microscopy (SEM) revealed that brown rice kernels had large contours of linear protuberances and depressions running lengthwise along the kernel surface. The protuberances were abraded successively during milling, but varying amounts of material remained in the depressions. Light microscopy combined with the lipid-specific probes Nile Blue A or Sudan Black B demonstrated that the material in the depressions observed with SEM was lipid. Sections of whole, milled rice kernels, prepared using a modified sectioning technique and stained with Nile Blue A, showed that portions of the embryo remain after milling and that lipid is located on or near the surface of the kernel. Differences in quantity and distribution of residual lipid as milling duration increased were documented photographically to indicate the extent to which the bran and embryo components were removed during milling. This paper provides proof of concept that residual lipid is a robust measure of the degree of milling.

Nanoemulsions prepared by a low-energy emulsification method applied to edible films.

Catastrophic phase inversion (CPI) was used as a low-energy emulsification method to prepare oil-in-water (O/W) nanoemulsions in a lipid (Acetem)/water/nonionic surfactant (Tween 60) system. CPIs in which water-in-oil emulsions (W/O) are transformed into oil-in-water emulsions (O/W) were induced by changes in the phase ratio. Dynamic phase inversion emulsification was achieved by slowly increasing the water volume fraction (fw) to obtain O/W emulsions from water in oil emulsions. Composition and processing variables were optimized to minimize droplet size and polydispersity index (PdI). It was found that addition of the continuous phase to the dispersed phase following the standard CPI procedure resulted in the formation of oil droplets with diameters of 100-200 nm. Droplet size distribution during CPI and emulsification time depended on stirring speed and surfactant concentration. Droplet sizes in the inverted emulsions were compared to those obtained by direct emulsification: The process time to reach droplet sizes of around 100 nm was reduced by 12 times by using CPI emulsification. The Acetem/water nanoemulsion was also used as a carrier to incorporate oregano and cinnamon essential oils into soy protein edible films. The resulting composite films containing oregano oil showed better moisture barrier and mechanical properties compared to soy protein films.

Composite edible films based on hydroxypropyl methylcellulose reinforced with microcrystalline cellulose nanoparticles.

It has been stated that hydroxypropyl methyl cellulose (HPMC) based films have promising applications in the food industry because of their environmental appeal, low cost, flexibility and transparency. Nevertheless, their mechanical and moisture barrier properties should be improved. The aim of this work was to enhance these properties by reinforcing the films with microcrystalline cellulose (MCC) at the nano scale level. Three sizes of MCC nanoparticles were incorporated into HPMC edible films at different concentrations. Identical MCC nanoparticles were lipid coated (LC) prior to casting into HPMC/LC-MCC composite films. The films were examined for mechanical and moisture barrier properties verifying how the addition of cellulose nanoparticles affected the water affinities (water adsorption/desorption isotherms) and the diffusion coefficients. The expected reinforcing effect of the MCC was observed: HPMC/MCC and HPMC/LC-MCC films showed up to 53% and 48% increase, respectively, in tensile strength values in comparison with unfilled HPMC films. Furthermore, addition of unmodified MCC nanoparticles reduced the moisture permeability up to 40% and use of LC-MCC reduced this value up to 50%. Water vapor permeability was mainly influenced by the differences in water solubility of different composite films since, in spite of the increase in water diffusivity values with the incorporation of MCC to HPMC films, better moisture barrier properties were achieved for HPMC/MCC and HPMC/LC-MCC composite films than for HPMC films.

Effect of fiber treatments on tensile and thermal properties of starch/ethylene vinyl alcohol copolymers/coir biocomposites.

Coir fibers received three treatments, namely washing with water, alkali treatment (mercerization) and bleaching. Treated fibers were incorporated in starch/ethylene vinyl alcohol copolymers (EVOH) blends. Mechanical and thermal properties of starch/EVOH/coir biocomposites were evaluated. Fiber morphology and the fiber/matrix interface were further characterized by scanning electron microscopy (SEM). All treatments produced surface modifications and improved the thermal stability of the fibers and consequently of the composites. The best results were obtained for mercerized fibers where the tensile strength was increased by about 53% as compared to the composites with untreated fibers, and about 33.3% as compared to the composites without fibers. The mercerization improved fiber-matrix adhesion, allowing an efficient stress transfer from the matrix to the fibers. The increased adhesion between fiber and matrix was also observed by SEM. Treatment with water also improved values of Young's modulus which were increased by about 75% as compared to the blends without the fibers. Thus, starch/EVOH blends reinforced with the treated fibers exhibited superior properties than neat starch/EVOH.

Nanocomposite edible films from mango puree reinforced with cellulose nanofibers.

Cellulose nanoreinforcements have been used to improve mechanical and barrier properties of biopolymers, whose performance is usually poor when compared to those of synthetic polymers. Nanocomposite edible films have been developed by adding cellulose nanofibers (CNF) in different concentrations (up to 36 g/100 g) as nanoreinforcement to mango puree based edible films. The effect of CNF was studied in terms of tensile properties, water vapor permeability, and glass transition temperature (T(g)) of the nanocomposite films. CNF were effective in increasing tensile strength, and its effect on Young's modulus was even more noticeable, especially at higher concentrations, suggesting the formation of a fibrillar network within the matrix. The addition of CNF was also effective to improve water vapor barrier of the films. Its influence on T(g) was small but significant. The study demonstrated that the properties of mango puree edible films can be significantly improved through CNF reinforcement.

Isolation and characterization of a cold-active xylanase enzyme from Flavobacterium sp.

Xylan is the major component of hemicellulose, and xylan should be fully utilized to improve the efficiencies of a biobased economy. There are a variety of industrial reaction conditions in which an active xylanase enzyme would be desired. As a result, xylanase enzymes with different activity profiles are of great interest. We isolated a xylanase gene (xyn10) from a Flavobacterium sp. whose sequence suggests that it is a glycosyl hydrolase family 10 member. The enzyme has a temperature optimum of 30 degrees C, is active at cold temperatures, and is thermolabile. The enzyme has an apparent Km of 1.8 mg/ml and kcat of 100 sec-1 for beechwood xylan, attacks highly branched native xylan substrates, and does not have activity against glucans.

An episomal expression vector for screening mutant gene libraries in Pichia pastoris.

Screening mutant gene libraries for isolating improved enzyme variants is a powerful technique that benefits from effective and reliable biological expression systems. Pichia pastoris is a very useful organism to express proteins that are inactive in other hosts such as Escherichia coli and Saccharomyces cerevisiae. However, most P. pastoris expression plasmids are designed to integrate into the host chromosome and hence are not as amenable to high-throughput screening projects. We have designed a P. pastoris expression vector, pBGP1, incorporating an autonomous replication sequence that allows the plasmid to exist as an episomal element. This vector contains the alpha-factor signal sequence to direct secretion of the mutant enzymes. Expression of the genes is driven by the constitutive GAP promoter, thus eliminating the need for timed or cell density-specific inductions. The pBGP1 plasmid was used to screen a xylanase gene library to isolate higher activity mutants.