Dimethylsulphopropionate (DMSP) and proline from the surface of the brown alga Fucus vesiculosus inhibit bacterial attachment

It was demonstrated previously that polar and non-polar surface extracts of the brown alga Fucus vesiculosus collected during winter from the Kiel Bight (Germany) inhibited bacterial attachment at natural concentrations. The present study describes the bioassay-guided identification of the active metabolites from the polar fraction. Chromatographic separation on a size-exclusion liquid chromatography column and bioassays identified an active fraction that was further investigated using nuclear magnetic resonance spectroscopy and mass spectrometry. This fraction contained the metabolites dimethylsulphopropionate (DMSP), proline and alanine. DMSP and proline caused the anti-attachment activity. The metabolites were further quantified on the algal surface together with its associated boundary layer. DMSP and proline were detected in the range 0.12–1.08 ng cm^?2 and 0.09–0.59 ng cm^?2, respectively. These metabolites were tested in the concentration range from 0.1 to 1000 ng cm^?2 against the attachment of five bacterial strains isolated from algae and sediment co-occurring with F. vesiculosus. The surface concentrations for 50% inhibition of attachment of these strains were always <0.38 ng cm^?2 for DMSP and in four cases <0.1 ng cm^?2 for proline, while one strain required 1.66 ng cm^?2 of proline for 50% inhibition. Two further bacterial strains that had been directly isolated from F. vesiculosus were also tested, but proved to be the least sensitive. This study shows that DMSP and proline have an ecologically relevant role as surface inhibitors against bacterial attachment on F. vesiculosus.     

Listeria monocytogenes Can Form Biofilms in Tap Water and Enter Into the Viable but Non-Cultivable State

Listeria monocytogenes is a foodborne pathogen that can be transmitted through contaminated raw food or by ready-to-eat products that have been in contact with contaminated surfaces. Tap water (TW) is used to wash produce, as a processed food constituent and to wash processing surfaces and floors. The main aim of this work was to investigate the formation and survival of L. monocytogenes biofilms on stainless steel (SS) coupons in TW at 4, 22, 30 and 37 °C. For that, coupons with biofilm were visualised in situ while other coupons were scraped to quantify total cells by SYTO 9, cultivable numbers by plating onto brain heart infusion agar and viable numbers by the direct viable count method. Results showed that L. monocytogenes can form biofilms on SS surfaces in TW at any temperature, including at 4 °C. The number of total cells was similar for all the conditions tested while cultivable numbers varied between the level of detection (<8.3 CFU cm^?2) and 3.5?×?10⁵ CFU cm^?2, meaning between 7.0?×?10⁴ and 1.1?×?10⁷ cells cm^?2 have entered the viable but non-cultivable (VBNC) state. This work clearly demonstrates that L. monocytogenes can form biofilms in TW and that sessile cells can remain viable and cultivable in some conditions for at least the 48 h investigated. On the other hand, VBNC adaptation suggests that the pathogen can remain undetectable using traditional culture recovery techniques, which may give a false indication of processing surface hygiene status, leading to potential cross-contamination of food products.     

Dependence of the initial adhesion of biofilm forming Pseudomonas putida mt2 on physico-chemical material properties

Bacterial adhesion is strongly dependent on the physico-chemical properties of materials and plays a fundamental role in the development of a growing biofilm. Selected materials were characterized with respect to their physico-chemical surface properties. The different materials, glass and several polymer foils, showed a stepwise range of surface tensions (γ_s) between 10.3 and 44.7 mN m^?1. Measured zeta potential values were in the range between ?74.8 and ?28.3 mV. The initial bacterial adhesion parameter q _max was found to vary between 6.6 × 10⁶ and 28.1 × 10⁶ cm^?2. By correlation of the initial adhesions kinetic parameters with the surface tension data, the optimal conditions for the immobilization of Pseudomonas putida mt2 were found to be at a surface tension of 24.7 mN m^?1. Both higher and lower surface tensions lead to a smaller number of adherent cells per unit surface area. Higher energy surfaces, commonly termed hydrophilic, could constrain bacterial adhesion because of their more highly ordered water structure (exclusion zone) close to the surface. At low energy surfaces, commonly referred to as hydrophobic, cell adhesion is inhibited due to a thin, less dense zone (depletion layer or clathrate structure) close to the surface. Correlation of q _max with zeta potential results in a linear relationship. Since P. putida carries weak negative charges, a measurable repulsive effect can be assumed on negative surfaces.     

The detection of food soils on stainless steel using energy dispersive X-ray and Fourier transform infrared spectroscopy

Organic soiling is a major issue in the food processing industries, causing a range of biofouling and microbiological problems. Energy dispersive X-ray (EDX) and Fourier transform infra red spectroscopy (FT-IR) were used to quantify and determine the biochemical groups of food soils on stainless steel surfaces. EDX quantified organic material on surfaces where oily based residues predominated, but was limited in its usefulness since other food soils were difficult to detect. FT-IR provided spectral ‘fingerprints’ for each of the soils tested. Key soiling components were associated with specific peaks, viz. oils at 3025 cm^?1–3011 cm^?1, proteins at 1698 cm^?1–1636 cm^?1 and carbohydrates at 1658 cm^?1–1596 cm^?1, 783 cm^?1–742 cm^?1. High concentrations of some soils (10%) were needed for detection by both EDX and FT-IR. The two techniques may be of use for quantifying and identifying specific recalcitrant soils on surfaces to improve cleaning and hygiene regimes.     

Biofilm formation by the yeast Rhodotorula mucilaginosa: process,repeatability and cell attachment in a continuous biofilm reactor

Yeast biofilms contribute to quality impairment of industrial processes and also play an important role in clinical infections. Little is known about biofilm formation and their treatment. The aim of this study was to establish a multi-layer yeast biofilm model using a modified 3.7 l bench-top bioreactor operated in continuous mode (D = 0.12 h^?1). The repeatability of biofilm formation was tested by comparing five bioprocesses with Rhodotorula mucilaginosa, a strain isolated from washing machines. The amount of biofilm formed after 6 days post inoculation was 83 μg cm^?2 protein, 197 μg cm^?2 polysaccharide and 6.9 × 10⁶ CFU cm^?2 on smooth polypropylene surfaces. Roughening the surface doubled the amount of biofilm but also increased its spatial variability. Plasma modification of polypropylene significantly reduced the hydrophobicity but did not enhance cell attachment. The biofilm formed on polypropylene coupons could be used for sanitation studies.     

TCNE-modified graphene as an adsorbent for N<Subscript>2</Subscript>O molecule: a DFT study

Adsorption behavior of nitrous oxide (N₂O) on pristine graphene (PG) and tetracyanoethylene (TCNE) modified PG surfaces is investigated using density functional theory. A number of initial adsorbate geometries are considered on both surfaces and the most stable ones are chosen upon calculation of the adsorption energies (E_ads). N₂O is found to adsorb in a weakly exoergic process (E_ads?～??3.18 kJ mol^?1) at the equilibrium distance of 3.52 Å on the PG surface. N₂O adsorption can be greatly enhanced with the presence of a TCNE molecule (E_ads?=??87.00 kJ mol^?1). Mulliken charge analysis confirms that adsorption of N₂O is not accompanied by distinct charge transfer from the surfaces to the molecule (? 0.001 │e│ for each case). Moreover, on the basis of calculated changes in the HOMO/LUMO energy gap, it is found that electronic properties of PG and TCNE modified PG are not sensitive toward adsorption of N₂O, indicating that both surfaces are not good enough to introduce as an N₂O detector. However, the considerable amount of E_ads in TCNE modified PG can be a guide to the design of graphene-based adsorbents for N₂O capture.     

Culture and exopolysaccharide production from sugarcane molasses by Gordonia polyisoprenivorans CCT 7137, isolated from contaminated groundwater in Brazil

Gordonia polyisoprenivorans CCT 7137 was isolated from groundwater contaminated with leachate in an old controlled landfill (São Paulo, Brazil), and cultured in GYM medium at different concentrations of sugarcane molasses (2%, 6%, and 10%). The strain growth was analyzed by monitoring the viable cell counts (c.f.u. mL^?1) and optical density and EPS production was evaluated at the end of the exponential phase and 24 h after it. The analysis of the viable cell counts showed that the medium that most favored bacterial growth was not the one that favored EPS production. The control medium (GYM) was the one that most favored the strain growth, at the maximum specific growth rate of 0.232 h^?1. Differences in bacterial growth when cultured at three different concentrations of molasses were not observed. Production of EPS, in all culture media used, began during the exponential phase and continued during the growth stationary phase. The highest total EPS production, after 24 h of stationary phase, was observed in 6% molasses medium (172.86 g L^?1) and 10% (139.47 g L^?1) and the specific total EPS production was higher in 10% molasses medium (39.03 × 10^?11 g c.f.u.^?1). After the exponential phase, in 2%, 6%, and 10% molasses media, a higher percentage of free exopolysaccharides (EPS) was observed, representing 88.4%, 62.4%, and 64.2% of the total, respectively. A different result was observed in pattern medium, which presented EPS made up of higher percentage of capsular EPS (66.4% of the total). This work is the first study on EPS production by G. polyisoprenivorans strain in GYM medium and in medium utilizing sugarcane molasses as the sole nutrient source and suggests its potential use for EPS production by G. polyisoprenivorans CCT 7137 aiming at application in biotechnological processes.     

Optimization and quality assessment of adventitious roots culture in Panax quinquefolium L.

In this study, adventitious roots of Panax quinquefolium L. have been successfully established. The highest induction rate of roots was obtained in MS medium containing 3 mg L^?1 IBA after 4 weeks of culture. The culture conditions of adventitious root were optimized and evaluated with response surface methodology. The best culture conditions for root growth seemed to be 0.75 salt strength MS medium, 4.70 g L^?1 inoculum size and 40 days of culture. The active component contents of adventitious roots were compared with those of native roots. The total saponins content was found to be 16.28 mg g^?1 in native root and 4.64 mg g^?1 in adventitious root. The polysaccharide content of the adventitious root was 1.5 times higher than that in the native P. quinquefolium (30.54 vs. 20.28 mg g^?1).     

A cost-effective method to quantify biological surface sediment reworking

We propose a simple and inexpensive method to determine the rate and pattern of surface sediment reworking by benthic organisms. Unlike many existing methods commonly used in bioturbation studies, which usually require sediment sampling, our approach is fully non-destructive and is well suited for investigating non-cohesive fine sediments in streams and rivers. Optical tracer (e.g. luminophores or coloured sand) disappearance or appearance is assessed through time based on optical quantification of surfaces occupied by tracers. Data are used to calculate surface sediment reworking (SSR) coefficients depicting bioturbation intensities. Using this method, we evaluated reworking activity of stream organisms (three benthic invertebrates and a fish) in laboratory microcosms mimicking pool habitats or directly in the field within arenas set in depositional zones. Our method was sensitive enough to measure SSR as low as 0.2 cm² day^?1, such as triggered by intermediate density (774 m^?2) of Gammarus fossarum (Amphipoda) in microcosms. In contrast, complex invertebrate community in the field and a fish (Barbatula barabatula) in laboratory microcosms were found to yield to excessively high SSR (>60 cm² day^?1). Lastly, we suggest that images acquired during experiments can be used for qualitative evaluation of species-specific effects on sediment distribution.     

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) co-polymer by the diazotrophic cyanobacterium Aulosira fertilissima CCC 444

Accumulation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate), P(3HB-co-3HV), a well-known co-polymer of polyhydroxyalkanoates family, was investigated in a N₂-fixing cyanobacterium, Aulosira fertilissima CCC 444, in presence of propionate and valerate in the culture medium. The most significant rise in P(3HB-co-3HV) co-polymer content up to 77 % of dry cell weight was recorded under 0.5 % fructose?+?0.4 % valerate supplementation depicting a productivity of 38 mg L^?1 day^?1, which was further increased by 2.5-fold, i.e., up to 95 mg L^?1 day^?1 under P deficiency. Surface analysis revealed a regular and smooth surface for P(3HB-co-3HV) co-polymer, against rugged and porous surface of the homopolymer of poly-β-hydroxybutyrate. X-ray diffraction showed semi-crystalline nature of the P(3HB-co-3HV) co-polymer. The thermal and mechanical properties of the co-polymer are comparable with the chemoheterotrophic bacterial polymers, thus opens up possibilities of using cyanobacterial PHAs in various fields.     

Theoretical description of the magnetic properties of μ3-hydroxo bridged trinuclear copper(II) complexes

A theoretical study of the magnetic properties, using density functional theory, of a family of trinuclear μ₃-OH copper(II) complexes reported in the literature is presented. The reported X-ray crystal structures of [Cu₃(μ₃-OH)(aat)₃(H₂O)₃](NO₃)₂·H₂O (HUKDUM), where aat: 3-acetylamine-1,2,4-triazole; [Cu₃(μ₃-OH)(aaat)₃(H₂SO₄)(HSO₄)(H₂O)] (HUKDOG), where aaat: 3-acetylamine-5-amine-1,2,4-triazole; [Cu₃(μ₃-OH)(PhPyCNO)₃(tchlphac)₂] (HOHQUR), where PhPyCNO: phenyl 2-pyridyl-ketoxime and tchlphac: acid 2,4,5-trichlorophenoxyacetic; [Cu₃(μ₃-OH)(PhPyCNO)₃(NO₃)₂(CH₃OH)] (ILEGEM); [Cu₃(μ₃-OH)(pz)₃(Hpz)₃(ClO₄)₂] (QOPJIP), where Hpz?=?pyrazole; [Cu₃(μ₃-OH)(pz)₃(Hpz)(Me₃CCOO)₂]?2Me₃CCOOH (DEFSEN) and [Cu₃(μ₃-OH)(8-amino-4-methyl-5-azaoct-3-en-2-one)₃][CuI₃] (RITXUO), were used in the calculations. The magnetic exchange constants were calculated using the broken-symmetry approach. The calculated J values are for HUKDUM J₁?=??68.6 cm^?1, J₂?=??69.9 cm^?1, J₃?=??70.4 cm^?1; for HUKDOG, J₁?=??73.5 cm^?1, J₂?=??58.9 cm^?1, J₃?=??62.1 cm^?1; for HOHQUR J₁?=??128.3 cm^?1, J₂?=??134.1 cm^?1, J₃?=??120.4 cm^?1; for ILEGEM J₁?=??151.6 cm^?1, J₂?=??173.9 cm^?1, J₃?=??186.9 cm^?1; for QOPJIP J₁?=??118.3 cm^?1, J₂?=??106.0 cm^?1, J₃?=??120.6 cm^?1; for DEFSEN J₁?=??74.9 cm^?1, J₂?=??64.0 cm^?1, J₃?=??57.7 cm^?1 and for RITXUO J₁?=??10.9 cm^?1, J₂?=?+14.3 cm^?1, J₃?=??35.4 cm^?1. The Kahn-Briat model was used to correlate the calculated magnetic properties with the overlap of the magnetic orbitals. Spin density surfaces show that the delocalization mechanism is predominant in all the studied compounds.

The invasive Sorghum halepense harbors endophytic N2-fixing bacteria and alters soil biogeochemistry

Exotic plants invading new habitats frequently initiate broad changes in ecosystem functioning. Sorghum halepense is an invasive grass capable of growing in nitrogen (N)-poor tallgrass prairie soils that creates near monocultures in once phylogenetically diverse-communities. The biogeochemistry of soils invaded by S. halepense was compared to that of un-invaded native prairie soils. Invaded soils contained two to four times greater concentrations of alkaline metals, micronutrients, and essential plant nutrients than native prairie soils. The notable exception was Ca⁺², which was always significantly lower in invaded soils. The N-content of S. halepense above-ground biomass was 6.4 mg g^?1 (320 mg N plant^?1) and suggested a supplemental N source supporting plant growth. Altered soil biogeochemistry in invaded areas coupled with high above-ground biomass in N-poor soils suggested N₂-fixing activity associated with S. halepense. Nitrogenase activity of plant tissues indicated that N₂-fixation was occurring in, and largely restricted to, S. halepense rhizomes and roots. A culture approach was used to isolate these N₂-fixing bacteria from plant tissues, and 16S rRNA gene sequencing was used to identify these bacterial isolates. Nitrogenase activity of bacterial isolates indicated several were capable of N₂-fixation. In addition to N₂-fixation, other roles involved in promoting plant growth, namely mobilizing phosphorus and iron chelation, are known for closest matching relatives of the bacterial isolates identified in this work. Our results indicate that these plant growth-promoting bacteria may enhance the ability of S. halepense to invade and persist by altering fundamental ecosystem properties via significant changes in soil biogeochemistry.     

Differences of skin morphology in Bos indicus,Bos taurus,and their crossbreds

Cutaneous evaporation is the main avenue by which cattle dissipate heat via the involvement of sweat glands and other skin components. The difference in skin morphology between B. indicus and B. taurus has been recognized, as well as differences in their ability to tolerate heat. The objective of this study was to compare skin morphology between B. indicus, B. taurus, and their crossbreds. Skin samples of Sahiwal (B. indicus) (n?=?10, reddish brown skin) and Holstein Friesian (HF) (B. taurus) (n?=?10, black and white skin) and crossbred of HF75% (n?=?10, black and white skin) and HF87.5 % (n?=?10, black and white skin) were biopsied for histological study, followed by measurement of skin components. The results indicated that breed significantly affected sweat gland morphology. The shape of the sweat gland, as indicated by the ratio of length/diameter, in Sahiwal was baggier in shape compared to HF (5.99 and 9.52) while values for crossbreds were intermediate (7.82, 8.45). The density and volume of sweat glands in Sahiwal (1,058 glands/cm²; 1.60 μ³?×?10^?6) were higher than in HF (920 glands/cm²; 0.51 μ³x10^?6) and crossbreds, both HF 75 % (709 glands/cm²; 0.68 μ³?×?10^?6) and HF 87.5 % (691 glands/cm²; 0.61 μ³?×?10^?6) respectively. However, capillary surface area was greater for HF (2.07 cm²) compared to Sahiwal (1.79 cm²); accordingly, the lower genetic fraction of HF in crossbred cattle showed less capillary surface area (1.83 and 1.9 cm² for HF75% and HF87.5 %) (P?<?0.01). Nerve density was not significantly different between Sahiwal and HF but was higher in the crossbred (P?<?0.01) cattle. Moreover, the effect of skin color (black and white) was evaluated and it was found that there was an interaction (P?<?0.01) between breed and skin color on the skin components. This study reveals that there are differences in skin morphology among B. indicus, B. taurus and their crossbreds, with these differences being more or less related to the genetic fraction of HF. This may imply that capability for cutaneous evaporative heat loss and tolerance to heat in crossbred cattle could be related to skin morphology.     

The influence of goethite and gibbsite on soluble nutrient dynamics and microbial community composition

Iron and aluminum (oxyhydr)oxides are ubiquitous in the soil environment and have the potential to strongly affect the properties of dissolved organic matter. We examined the effect of oxide surfaces on soluble nutrient dynamics and microbial community composition using an incubation of forest floor material in the presence of (1) goethite and quartz, (2) gibbsite and quartz, and (3) quartz surfaces. Forest floor material was incubated over a period of 154 days. Aqueous extracts of the incubations were harvested on days 5, 10, 20, 30, 60, 90, and 154, and concentrations of P, N, PO₄ ^3?, NO₂ ^?, NO₃ ^?, and organic C were measured in the solutions. Microbial community composition was examined through pyrosequencing of bacterial and fungal small subunit ribosomal RNA genes on selected dates throughout the incubation. Results indicated that oxide surfaces exerted strong control on soluble nutrient dynamics and on the composition of the decomposer microbial community, while possibly having a small impact on system-level respiration. Goethite and gibbsite surfaces showed preferential adsorption of P-containing and high molar mass organic solutes, but not of N-containing compounds. On average, organic C concentrations were significantly lower in water extractable organic matter (WEOM) solutions from oxide treatments than from the control treatment (P = 0.0037). Microbial community composition varied both among treatments and with increasing time of incubation. Variation in bacterial and fungal community composition exhibited strong-to-moderate correlation with length of incubation, and several WEOM physiochemical characteristics including apparent (weight averaged) molar mass, pH and electrical conductivity. Additionally, variation in bacterial community composition among treatments was correlated with total P (r = 0.60, P < 0.0001), PO₄ ^3? (r = 0.79, P < 0.0001), and organic C (r = 0.36, P = 0.015) concentrations; while variation in fungal communities was correlated with organic C concentrations (r = ?0.48, P = 0.0008) but not with phosphorus concentrations. The relatively small impact of oxide surfaces on system-level microbial respiration of organic matter despite their significant effects on microbial community composition and WEOM dynamics lends additional support to the theory of microbial functional redundancy.     

Epiphytism in a subtidal natural bed of Gracilaria gracilis of southwestern Atlantic coast (Chubut, Argentina)

The diversity of epiphytes, their temporal abundance variation and the anatomical structure of host–epiphyte interfaces were studied in the agarophyte Gracilaria gracilis from a natural bed in Bahía Bustamante, Chubut Province, Argentina. Twenty-nine algal species were recorded as epiphytes during 2 years of monthly sampling (March 2006–February 2008). Total epiphyte density ranged between 0.037 ind. cm^?2 (November 2006) and 39.37 ind. cm^?2 (April 2007), with higher density values throughout the second sampling year. Ceramiales species were the most abundant epiphytes. The density of Ceramium rubrum ranged from 0.09 ind. cm^?2 (52 % of the total amount) in September 2006 to 17.4 ind. cm^?2 (44.18 % of the total amount) in April 2007. Epiphyte infection was more dependent on spore recruitment and sporeling development, especially on thalli derived from fragmentation, than on seasonal environmental variations. The different infections were analysed, taking into account the epiphytic attachment strength and invasiveness and the degree of damage inflicted on the host. Calothrix confervicola was one of the most abundant species. This epiphyte, weakly attached to the host surface, generated no host tissue damage. In contrast, C. rubrum, Polysiphonia abscissa and other Ceramiales were the species that caused more damage to the host because their rhizoids penetrated the cortical portion of the host thallus, sometimes reaching the medullary tissue. Some generalisations and characterisations of the different epiphyte groups in relation to their consequences to Gracilaria spp. are presented.     

Direct discrimination of different plant populations and study on temperature effects by Fourier transform infrared spectroscopy

Fourier transform infrared spectroscopy was used to characterise highland and lowland populations of Polygonum minus Huds. grown in different controlled environments. A thermal perturbation technique of two-dimensional correlation infrared spectroscopy (2D-IR) correlation spectra was applied to establish differences between the populations. The absorption peaks at 3,480 cm^?1 (hydroxyl group), 2,927 cm^?1 (methyl group), 1,623 cm^?1 (carbonyl group), and 1,068 cm^?1 (C–O group) were particularly powerful in separating the populations. These peaks, which indicate the presence of carbohydrate, terpenes, amide and flavonoids were more intense for the highland populations than lowland populations, and increased in environments with a higher temperature. Wavenumbers (1,634, 669 cm^?1) and (1,634, 1,555 cm^?1) in the 2D-IR correlation spectra provided fingerprint signals to differentiate plants grown at different temperatures. This study demonstrates that IR fingerprinting, which combines mid-IR spectra and 2D-IR correlation spectra, can directly discriminate different populations of P. minus and the effects of temperature.     

Edaphic constraints on seed germination and emergence of three <Emphasis Type="Italic">Acacia</Emphasis> species for dryland restoration in Saudi Arabia

In situ edaphic factors affecting seed germination and seedling emergence of three framework species of Acacia were investigated with the intent of developing fundamental and scalable restoration capacity for Arabian dryland restoration. Direct seeding represents the most efficient means to restore vegetation at the landscape scale and this study provides insight into edaphic and ecological limitations, as well as effective protocols governing the use of native seeds for restoration in hyper-arid environments. The study was conducted in extant Acacia woodland habitat on conserved land (Thumamah Nature Park) in close proximity to Riyadh, Saudi Arabia. Broad-scale direct seeding using un- and pretreated Acacia gerrardii, A. tortilis, and A. ehrenbergiana seed, and two seed burial depths were implemented across three sites with distinct soil surface characteristics. Eight weeks post-sowing, random samples for each species × seed treatment × burial depth combination were excavated, sieved, and categorized as follows: failed to germinate, germinated but died prior to emerging, or successfully emerged. We show that germination and emergence of Acacia gerrardii, A. tortilis, and A. ehrenbergiana were driven by a three-way interaction among species, site, and seed burial depth. Treating seed with the signaling compound Moddus did not have a definitive effect, positive or negative, on any of the species investigated. Acacia gerrardii was the only species that exhibited widespread emergence, though emergence was not consistent across sites or burial depths. Germination was highest in disturbed soil (up to 69% for A. gerrardii), but very few (<2%) successfully emerged; a greater proportion of germinants in sandy soil emerged (up to 44% for A. gerrardii) even though the overall germination was less. Though species-dependent, a 2-cm sowing depth was most effective in sand; while in disturbed soil, sowing depths of 1 and 2 cm were comparable; and no germination was observed in gravelly clay soil. Sandy soil exhibited rapid water infiltration (107.6 mm min^?1), and post-sowing surface crusting was a non-factor (0.44 kg cm^?2). Disturbed soil exhibited moderate water infiltration (1.46 mm min^?1) and post-sowing surface crusting was double that of sand (0.88 kg cm^?2) and restrictive on seedling emergence. Gravelly clay exhibited extremely poor water infiltration (0.12 mm min^?1), and surface crusting was severe (4.49 kg cm^?2) and an order of magnitude greater than sand. The medium-coarse sand fraction, a key driver of the observed soil surface processes, was greatest in sand (55%) and significantly less and uniform in the disturbed (22%) and gravelly clay (22%) soils. Our findings demonstrate that soil surface characteristics and associated processes can dictate ecological processes at depths as shallow as 1–2 cm, and that soil crusts that slow water infiltration and impede seedling emergence rapidly reconstitute after disturbance; both are important considerations for restoring dryland vegetation.     

Non-native liana, <Emphasis Type="Italic">Euonymus fortunei</Emphasis>, associated with increased soil nutrients,unique bacterial communities,and faster decomposition rate

Invasive plants have wide-ranging impacts on native systems including reducing native plant richness and altering soil chemistry, microbes, and nutrient cycling. Increasingly, these effects are found to linger long after removal of the invader. We examined how soil chemistry, bacterial communities, and litter decomposition varied with cover of Euonymus fortunei, an invasive evergreen liana, in two central Kentucky deciduous forests. In one forest, E. fortunei invaded in the late 1990s but invasion remained patchy and we paired invaded and uninvaded plots to examine the associations between E. fortunei cover and our response variables. In the second forest, E. fortunei had completely invaded the forest by 2005; areas where it had been selectively removed by 2010 were paired with an adjacent invaded plot. Where E. fortunei had patchily invaded, E. fortunei patches had up to 3.5× nitrogen, 2.7× carbon, and 1.9× more labile glomalin in soils than uninvaded plots, whereas there were no differences in soil characteristics between invaded and removal plots. In the patchily invaded forest, bacterial community composition varied among invaded and non-invaded plots, whereas bacterial communities did not vary among invaded and removal plots. Finally, E. fortunei leaf litter decomposed faster (k = 4.91 year^?1) than the native liana (k = 3.77 year^?1), Vitis vulpina; decomposition of both E. fortunei and V. vulpina was faster in invaded (k = 7.10 year^?1) than removal plots (k = 4.77 year^?1). Our findings suggest that E. fortunei invasion increases the rate of leaf litter decomposition via high-quality litter, alters the decomposition environment, and shifts in the soil biotic communities associated with a dense mat of wintercreeper. Land managers with limited resources should target the densest mats for the greatest restoration potential and remove wintercreeper patches before they establish dense mats.     

Enhanced efficacy of nitrifying biomass by modified PVA_SB entrapment technique

In this study, we developed a novel technique for preparing polyvinyl alcohol (PVA) hydrogel as an immobilizing matrix by the addition of sodium bicarbonate. This resulted in an increase in the specific surface area of PVA_sodium bicarbonate (PVA_SB) hydrogel beads to 65.23 m² g^?1 hydrogel beads, which was approximately 85 and 14 % higher than those of normal PVA and PVA_sodium alginate (PVA_SA) hydrogel beads, respectively. The D _e value of PVA_SB hydrogel beads was calculated as 7.49 × 10^?4 cm² s^?1, which was similar to the D _e of PVA_SA hydrogel beads but nearly 38 % higher than that of the normal PVA hydrogel beads. After immobilization with nitrifying biomass, the oxygen uptake rate and the ammonium oxidation rate of nitrifying biomass entrapped in PVA_SB hydrogel beads were determined to be 19.53 mg O₂ g MLVSS^?1 h^?1 and 10.59 mg N g MLVSS^?1 h^?1, which were 49 and 43 % higher than those of normal PVA hydrogel beads, respectively. Scanning electron microscopy observation of the PVA_SB hydrogel beads demonstrated relatively higher specific surface area and revealed loose microstructure that was considered to provide large spaces for microbial growth. This kind of structure was also considered beneficial for reducing mass transfer resistance and increasing pollutant uptake.     

A new biopesticide from a local <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> var. <Emphasis Type="Italic">tenebrionis</Emphasis> (Xd3) against alder leaf beetle (Coleoptera: Chrysomelidae)

Use of chemical pesticides in agriculture harms humans, non-target organisms and environments, and causes increase resistance against chemicals. In order to develop an effective bio-pesticide against coleopterans, particularly against Agelastica alni (Coleoptera: Chrysomelidae) which is one of the serious pests of alder leaf and hazelnut, we tested the insecticidal effect of 21 Bacillus isolates against the larvae and adults of the pest. Bacillus thuringiensis var. tenebrionis-Xd3 (Btt-Xd3) showed the highest insecticidal effect based on screening tests. For toxin protein production and high sporulation of Xd3, the most suitable medium, pH and temperature conditions were determined as nutrient broth medium enriched with salts, pH 7 and 30?°C, respectively. Sporulated Btt-Xd3 in nutrient broth medium enriched with salts transferred to fermentation medium containing soybean flour, glucose and salts. After fermentation, the mixture was dried in a spray dryer, and spore count of the powder product was determined as 1.6?×?10¹⁰ c.f.u. g^?1. Moisture content, suspensibility and wettability of the formulation were determined as 8.3, 86% and 21 s, respectively. Lethal concentrations (LC₅₀) of formulated Btt-Xd3 were determined as 0.15?×?10⁵ c.f.u. ml^?1 for larvae at laboratory conditions. LC₅₀ values were also determined as 0.45?×?10⁶ c.f.u. ml^?1 at the field condition on larval stage. Our results showed that a new bio-pesticide developed from B. thuringiensis tenebrionis (Xd3) (Btt-Xd3) may be valuable as a biological control agent for coleopteran pests.