Purification and characterization of a Na+/K+ dependent alginate lyase from turban shell gut Vibrio sp. YKW-34

An alginate lyase with high specific enzyme activity was purified from Vibrio sp. YKW-34, which was newly isolated from turban shell gut. The alginate lyase was purified by in order of ion exchange, hydrophobic and gel filtration chromatographies to homogeneity with a recovery of 7% and a fold of 25. This alginate lyase was composed of a single polypeptide chain with molecular mass of 60 kDa and isoelectric point of 5.5–5.7. The optimal pH and temperature for alginate lyase activity were pH 7.0 and 40 °C, respectively. The alginate lyase was stable over pH 7.0–10.0 and at temperature below 50 °C. The alginate lyase had substrate specificity for both poly-guluronate and poly-mannuronate units. The k_cat/K_m value for alginate (heterotype) was 1.7 × 10⁶ s⁻¹ M⁻¹. The enzyme activity was completely lost by dialysis and restored by addition of Na⁺ or K⁺. The optimal activity exhibited in 0.1 M of Na⁺ or K⁺. This enzyme was resistant to denaturing reagents (SDS and urea), reducing reagents (β-mercaptoethanol and DTT) and chelating reagents (EGTA and EDTA).     

Soil microbial biomass influence on growth and biocontrol efficacy of Trichoderma harzianum

Hyphal growth and biocontrol efficacy of Trichoderma harzianum may depend on its interactions with biotic components of the soil environment. Effects of soil microbial biomass on growth and biocontrol efficacy of the green fluorescent protein transformant T. harzianum ThzID1-M3 were investigated using different levels of soil microbial biomass (153, 328, or 517 μg biomass carbon/g of dry soil). Hyphal growth of T. harzianum was significantly inhibited in soil containing 328 or 517 μg biomass carbon/g of dry soil compared with soil containing 153 μg biomass carbon/g. However, when ThzID1-M3 was added to soil as an alginate pellet formulation, recoverable populations of ThzID1-M3 varied, with the highest populations in soil containing 517 μg biomass carbon/g. When sclerotia of Sclerotinia sclerotiorum were added to soils (10 sclerotia per 150 g soil) with ThzID1-M3 (20 pellets per 150 g soil), colonization of sclerotia by ThzID1-M3 was significantly lower in the soil containing the highest level of biomass. Addition of alginate pellets of ThzID1-M3 to soils (10 pellets per 50 g) resulted in increased indigenous microbial populations (total fungi, bacteria, fluorescent Pseudomonas spp., and actinomycetes). Our results suggest that higher levels of microbial soil biomass result in increased interactions between introduced T. harzianum and soil microorganisms, and further that microbial competition in soil favors a shift from hyphal growth to sporulation in T. harzianum, potentially reducing its biocontrol efficacy.     

Actividad antifúngica de los enjuagues bucales frente a Candida albicans y Rhodotorula mucilaginosa: un estudio in vitro

BackgroundCandida albicans and Rhodotorula mucilaginosa are yeasts of clinical importance in the oral cavity. In immunocompromised patients they can cause some pathologies that must be controlled with antimicrobials.AimsTo evaluate and compare the antimicrobial efficacy of commercially available mouthrinses against strains of C. albicans and R. mucilaginosa.MethodsThe six mouthwashes studied in vitro were formulated (alone or in combination) with chlorhexidine (CHX) 0.12%, CHX 0.1%, CHX 0.05%, cetylpyridinium chloride (CPC) 0.075%, CPC 0.05%, and essential oils. Ten C. albicans and R. mucilaginosa isolates each were studied. The agar diffusion method (Mueller Hinton II), with incubation at 32 °C was used to evaluate the antifungal activity.ResultsThe results of this study indicate that mouthwashes with CHX 0.1%, CHX 0.12%, CHX 0.05% + CPC 0.05%, CHX 0.12% + CPC 0.05% and CPC 0.075% have an antifungal effect against C. albicans and R. mucilaginosa. CHX 0.1% led to the broadest inhibition zone for C. albicans and R. mucilaginosa (25.65 ± 2.39 mm and 40.05 ± 3.31 mm). Essential oils did not show any antifungal activity. Statistical analysis showed no statistical difference between mouth rinses CHX 0.1%, CHX 0.12% and CHX 0.12% + CPC 0.05% (p = 0.0001) against C. albicans and R. mucilaginosa.ConclusionsMouthwashes with CHX showed higher antifungal activity against C. albicans and R. mucilaginosa than other mouthwashes studied.     

Effect of high temperature on active oxygen species,senescence and photosynthetic properties in cucumber leaves

To gain a physiological understanding of the effects of high temperatures on cucumber (Cucumis sativus L.), we subjected seedlings to heat treatment at daytime temperatures of 28 °C, 32 °C, and 36 °C for 7 h a day for 30 days. The amount of active oxygen species, indicators of senescence, and photosynthetic properties in the second and third leaves were determined at the start of temperature treatment and on the 15th and 30th days of treatment. The amount of active oxygen species superoxide in leaves was greatest in the high temperature zones on the 15th day of treatment, and the amount of hydrogen peroxide was greatest in the high temperature zones on both the 15th and 30th days of treatment. The reduction in the amount of protein and the increase in the amount of malondialdehyde, both indicators of senescence, were greatest in the high temperature zones on both the 15th and 30th days of treatment, and the amount of chlorophyll was lowest in the 36 °C zone on the 15th day, and lower in the high temperature zones on the 30th day. It is clear from these results that a large amount of active oxygen species is generated and accumulated in the leaves at high temperatures, and senescence is significantly accelerated. The photosynthetic properties of stomatal conductance, sub-stomatal CO₂ concentration, and transpiration rate were at the same level on both the 15th and 30th days of treatment in all three temperature treatment zones. No significant difference was seen in the net photosynthesis rate between the 28 °C and 32 °C zones, was lower in the 36 °C zone than the 32 °C zone on the 15th day, and lowest in the 36 °C zone on the 30th day. CO₂ intake and water absorption are only mildly affected by high temperatures, and the reduction in net photosynthesis rate due to the 36 °C high temperature stress suggests that the large amount of active oxygen species induces inhibition of photosynthesis and damage to the mechanism of photosynthesis.     

Dried alginate-entrapped enzymes (DALGEEs) and their application to the production of fructooligosaccharides

A modification of the classical calcium alginate enzyme entrapment technique is described aiming to overcome some of the limitations of the former gel-based biocatalysts. Dried alginate entrapped enzymes (DALGEEs) were obtained dehydrating calcium alginate gel beads containing entrapped enzymes. A fructosyltransferase from Aspergillus aculeatus, present in Pectinex Ultra SP-L, was entrapped using this technique. The resulting DALGEEs were successfully tested both operating batchwise and in a continuous fixed-bed reactor for fructooligosaccharides (FOS) synthesis from sucrose. Interestingly, DALGEEs did not re-swell upon incubation in concentrated (600 g/L) sucrose solutions, probably due to the lowered water activity (a_w) of such media. Confocal laser scanning microscopy of DALGEEs revealed that the enzyme molecules accumulated preferably in the shell of the particles. DALGEEs showed an approximately 30-fold higher volumetric activity (300 U/mL) compared with the calcium alginate gel beads. Moreover, a significant enhancement (40-fold) of the space-time-yield of fixed-bed bioreactors was observed when using DALGEEs as biocatalyst compared with gel beads (4030 g/day L of FOS vs. 103 g/day L). The operational stability of fixed-bed reactors packed with DALGEEs was extraordinary, providing a nearly constant FOS composition of the outlet during at least 700 h. It was also noticeable their resistance against microbial attack, even after long periods of storage at room temperature. The DALGEEs immobilisation strategy may also be useful for other biotransformations, in particular when they take place in low a_w media.     

Screening for hydroxynitrile lyase activity in non-commercialised plants

Hydroxynitrile lyases are used for the synthesis of enantiomerically pure cyanohydrins which are of great importance in the pharmaceutical and fine chemical industries. In this study, the hydroxynitrile lyase activity of 100 plants from 40 families was investigated, first by screening for cyanogenic activity, followed by a hydroxynitrile lyase activity assay. Of the 100 plants, four were found to be cyanogenic and exhibited specific hydroxynitrile lyase activity: Adenia sp. (0.44 U/mg), Adenia firingalavensis (2.88 U/mg), Adenia fruticosa (1.99 U/mg) and, Adenia pechuelii (2.35 U/mg), all from the family Passifloraceae. This is the first report of hydroxynitrile lyase activity in these plants.     

Examination of six commonly used laboratory fixatives in HAB monitoring programs for their use in quantitative PCR based on Taqman probe technology

Due to the need for more rapid and reliable detection, quantification and enumeration of harmful algal species the use of molecular methods are increasingly being used in monitoring and field studies. However, many studies often require sample fixation to allow for transportation before analyses are conducted. Here, we describe the effects of six fixatives (acidified Lugol's iodine with or without sodium thiosulphate, glutaraldehyde, paraformaldehyde (PFA), formalin and ethanol) on quantitative real-time polymerase chain reaction (qPCR) amplification with Taqman probes. We applied extracted total genomic DNA from four harmful algal species from Danish waters, representing three dinoflagellates (Alexandrium tamarense, Karenia mikimotoi, Karlodinium veneficum and a haptophyte (Prymnesium parvum). The C_q values generated on the qPCR amplification plot were compared to those of an unfixed sample that acted as a control. For all species positive amplifications were achieved from DNA templates from all preserved samples. However, amplification efficiencies between fixatives and species varied. Yet it was found that Lugol's iodine was the most ideal short-term fixative for enumeration of cells by qPCR as well as being the safest to handle. The effect of age on Lugol's iodine fixed samples was also addressed. Samples were fixed and stored at 5 °C in the dark and total genomic DNA extracted after 24 h, 72 h, 1 week, 2 weeks, 1 month and 2 months. Samples remained stable for 1 month for A. tamarense and K. veneficum and 2 months for K. mikimotoi and P. parvum.     

Temporal–spatial variation and source analysis of carbon and nitrogen in a tidal wetland of Luoyuan Bay

Carbon and nitrogen are important elements in biogeochemical studies of tidal wetlands. Three wetland zones in Luoyuan Bay in the Fujian province were chosen for this study; the Spartina alterniflora flat zone with Spartina alterniflora growing, the silt zone with no Spartina alterniflora growing and the Spartina alterniflora-silt flat zone – a transition zone between the two. The spatial and seasonal variations of total organic carbon (TOC), total nitrogen (TN), stable isotopes of organic material (δ¹³C, δ¹⁵N), C/N ratio, average particle size and sediment composition in surface and vertical sediments of different ecological zones were analyzed. Carbon and nitrogen accumulation and particle size effects in the different ecological zones were discussed and the indicators of δ¹³C and C/N ratios were also compared. TOC, TN, δ¹³C contents, C/N ratios, and average particle size varied within the ranges of 0.611–1.133%, 0.053–0.090%, ?22.60 to ?18.92‰, 12.3–15.7, and 6.4–8.7 μm, respectively. Sediments were mainly silt-sized. Besides δ¹⁵N values, the other parameters, such as TOC, TN, δ¹³C contents, C/N ratios, and average particle size showed an obvious zonal distribution in surface sediments. The distribution of TOC and TN contents reflected the distribution of Spartina alterniflora within the bay. The profile and seasonal variations of these parameters in different ecological zones indicated that variations in the Spartina alterniflora flat and transition zones were complex because of the effect of Spartina alterniflora. Vertical and seasonal variations were sampled in the silt flat area. The profile and seasonal variations of TOC, TN and δ¹³C were similar in the transition zone and the Spartina alterniflora flat zone. Seasonal concentrations of TOC, TN and δ¹³C decreased from autumn > spring > winter > summer. The seasonal variation of carbon and nitrogen in the sediments may be influenced by temperature, particle size, plankton and benthos. The particle size effect was significant in the surface sediments and profile sediments of the transition zone. However, other factors had a greater effect on the distributions of TOC and TN in the Spartina alterniflora flat and silt flat zones. C/N ratios in sediments of the Spartina alterniflora flat, transition zone and silt flat were close to or > 12, indicating that the organic material source was dominated by terrestrial inputs. However, δ¹³C values decreased from the Spartina alterniflora flat zone > transition zone > silt flat zone indicating that the organic material source was predominantly from marine inputs. Thus the indications from C/N ratios and δ¹³C were different. There was no clear relationship between C/N ratios and δ¹³C values and a better relationship between δ¹³C values and TOC concentrations suggested that δ¹³C values provided a better indication of the organic source. Limited amounts of organic material came from Spartina alterniflora. This study has provided basic data for researching biogeochemical processes of biogenic elements in tidal wetlands and vegetation restoration, and has also provided a reference for assessing and protecting the environment and ecological systems in wetlands.     

The historical dendroarchaeology of two log structures at the Wynnewood State Historic Site,Castalian Springs,Tennessee, USA

Tree rings of eastern red cedar (Juniperus virginiana L.) were examined from cores extracted from two log cabins located at the Wynnewood State Historic Site in Castalian Springs, Sumner County, Tennessee. One cabin was reportedly built by the first explorer in the area, Isaac Bledsoe, sometime between 1772 and his death in 1793. The second cabin was known as Spencer's Cabin after the first settler of the region, Thomas Sharp Spencer, who lived in the immediate vicinity from 1776 to 1779. The goal of this research was to determine the probable construction year(s) for both cabins and determine whether Bledsoe and Spencer did indeed build these structures. Forty-one cores were extracted from Bledsoe's Cabin, and 30 were used for crossdating and building a floating chronology using COFECHA. The Bledsoe's Cabin chronology was then statistically and graphically crossdated using the eastern red cedar reference tree-ring chronology (ITRDB #TN031) from Norris Dam, Tennessee. We found a statistically significant correlation (r = 0.42, t = 4.18, n = 85, p < 0.0001) between the Bledsoe's Cabin chronology and the reference chronology, anchoring the chronology between 1720 and 1804, with nearly all cores indicating tree harvesting between February and April 1805. Twenty-two cores were extracted from Spencer's Cabin, and 17 were used to build a floating chronology for the cabin. Again, we found a statistically significant correlation (r = 0.44, t = 4.85, n = 100, p < 0.00001) with the reference chronology, which anchored the Spencer's Cabin chronology between 1726 and 1825. All trees appear to have been harvested between February and August in 1826. Therefore, neither structure was built by its historical namesake. No known historical documents suggest who the potential builders were, although the property was owned between ca. 1797 and 1829 by General James Winchester. He and his family, however, never resided on the Wynnewood property because Winchester had built a large multi-room structure in nearby Gallatin, Tennessee, by 1802.     

The hydrolysate of barley straw containing inhibitors can be used to produce cephalosporin C by solvent extraction using ethyl acetate

When the solvent extraction of the hydrolysate from barley straw was performed using ethyl acetate (EA), the logarithm of the partition coefficient (log P) of the phenols and furans for EA was found to be more than 1.00, which means that more than 90% of the inhibitors were removed from the hydrolysate layer. Cephalosporin C (CPC) was produced from the hydrolysate of dilute acid pretreatment (DAP) by Acremonium chrysogenum M35. A. chrysogenum M35 was cultured using the hydrolysate and the amount of CPC produced was found to be 10.35 g/L at 144 h. Also, the dry cell weight was about 101.5 g/L at 120 h. The utilization of the hydrolysate for CPC production was effective and the solvent extraction method for the removal of inhibitory substances could contribute to the biorefinery process.     

Thermal analysis for differentiating between oleaginous and non-oleaginous microorganisms

The potential of thermal analysis for differentiating between oleaginous and non-oleaginous microorganisms was investigated using thermogravimetry (TG) and differential thermal analysis (DTA). The model oleaginous microorganisms used in the present study were the fungi, Mortierella alpina IFO32281 and Mortierella alliacea YN-15, the unicellular alga, Aurantiochytrium sp. CB 15-5, and the yeast, Rhodosporidium toruloides DMKU3-TK 16. Escherichia coli JM109, Rhodococcus opacus B-4, and Saccharomyces cerevisiae were used as the control non-oleaginous microorganisms. In simultaneous TG and DTA, the furnace temperature was linearly increased from 30 to 280 °C, decreased to 30 °C, linearly increased from 30 to 360 °C, and then isothermally held at 360 °C for 30 min. This two-step linear temperature program was effective in resolving overlapping exothermic peaks in the DTA curves in the temperature range from 280 to 360 °C. Heat evolved from a microbial sample was estimated from the area under the exothermic peak between 280 and 360 °C using indium as a standard material. There was a linear relationship between the exothermic heat and total lipid content of the tested microorganisms. Exothermic heat per dry sample mass (kJ/g) in the temperature range from 280 to 360 °C is a promising measure for differentiating between oleaginous and non-oleaginous microorganisms.     

Synthesis,characterization and swelling behaviors of sodium alginate-g-poly(acrylic acid)/sodium humate superabsorbent

A novel sodium alginate-g-poly(acrylic acid)/sodium humate superabsorbent was prepared by graft copolymerization with sodium alginate, acrylic acid and sodium humate in aqueous solution, using N,N’-methylenebisacrylamide as a crosslinker and ammonium persulfate as an initiator. The effects of crosslinker, sodium alginate and sodium humate content on water absorbency of the superabsorbent were studied. The swelling behavior in solutions of various pH and the swelling kinetics in saline solutions (5 mmol/L NaCl and CaCl₂) were also investigated. The results from IR analysis showed that both sodium alginate and sodium humate react with the acrylic acid monomer during the polymerization process. The introduction of sodium humate into the sodium alginate-g-poly(acrylic acid) system could enhance the water absorbency and the superabsorbent containing 10 wt% sodium humate acquired the highest water absorbency (1380 g/g in distilled water and 83 g/g in 0.9 wt% NaCl solution).     

Can surface electromyography improve surgery planning? Electromyographic assessment and intraoperative verification of the nerve bundle entry point location of the gracilis muscle

Purpose: To verify the precision of surface electromyography (sEMG) in locating the innervation zone of the gracilis muscle, by comparing the location of the IZ estimated by means of sEMG with in vivo location of the nerve bundle entry point in patients before graciloplasty procedure due to fecal incontinence. Methods: Nine patients who qualified for the graciloplasty procedure underwent sEMG on both gracilis muscle before their operations. During surgery the nerve bundle was identified by means of electrical stimulation. The distance between the proximal attachment and the nerve entry point into the muscle’s body was measured. Both measurements (sEMG and in vivo identification) were compared for each subject. Results: On average, the IZ was located 65.5 mm from the proximal attachment. The mean difference in location of the innervation zones in each individual was 10 ± 9.7 mm, maximal – 30 mm, the difference being statistically significant (p = 0.017). It was intraoperatively confirmed, that the nerve entered the muscle an average of 62 mm from the proximal attachment. The largest difference between the EMG IZ estimation and nerve bundle entry point was 5 mm (mean difference 2.8 mm, p = 0.767). Conclusion: Preoperative surface electromyography of both gracilis muscles is a safe, precise and reliable method of assessing the location of the innervation zones of the gracilis muscles. The asymmetry of the IZ location in left and right muscles may be important in context of technical aspects of the graciloplasty procedure.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Microbial contamination of traditional medicinal plants sold at the Faraday muthi market,Johannesburg, South Africa

The Faraday traditional healers' trading market is the hub of the medicinal plant trade in Johannesburg, South Africa. Modes of harvesting, transporting, storage and distribution of medicinal plants render them susceptible to microbial attack, and thereby make customers, especially patients with compromised immune systems, vulnerable to infections that could increase morbidity and mortality. This study evaluated the microbial contamination on five frequently used traditional medicinal plant species sold by traders in the Faraday market. Bacterial contamination was determined using serial macro dilutions, spread plate and streak plate techniques. Fifteen bacterial contaminants were identified, the most recurrent being Pantoea sp. and five strains of Bacillus spp. (non-pathogenic). There was little variation between contamination levels of the five different traders, and the mean CFU/g per species ranged from 3.03 × 10⁴ (Hypoxis sp.) to 4.22 × 10⁵ (Hydnora abyssinica). While there was no overall significant levels of contamination, the CFU counts for two plant species purchased from one specific trader (viz. H. abyssinica and Acacia xanthophloea) exceeded maximum acceptable contamination limits set by the World Health Organisation (i.e. ≤ 10⁵ to ≤ 10⁷ CFU/g). The levels of contamination varied greatly between the commercially available over the counter product and the plant samples investigated. The microbial types are predominantly opportunistic pathogens. The implementation of good processing practices therefore clearly influences the quality and safety of medicinal products, especially regarding microbial contamination. It is evident that policies and regulations need to be developed and implemented in order to address possible contamination by opportunistic pathogens.     

Effect of polymer/nanosilver composite packaging on long-term microbiological status of Iranian saffron (Crocus sativus L.)

Crocus sativus L. (saffron) is a valuable plant which is native to Iran. Saffron is the dried stigmata of the flowering part of the plant that is usually contaminated with different bacteria and fungi through production process. Antimicrobial properties of silver nanoparticles are well recognized. To survey the effects of nanosilver packaging on microbiological status of spiked, saffron samples over a six month period were chosen. Saffron samples from five regions of Khorasan province were purchased and de novo frequencies of microbial contaminants were determined using standard procedures. Totally 35 g of saffron was spiked with known numbers of four bacterial and two fungal species and packaged into one gram packets. The packaging materials consisted of polyethylene polymers containing 0, 400, 800, 1200 or 4000 ppm nanosilver (as Ag). Total and differential numbers of spiked microorganisms in the packaged saffrons were enumerated at initial and at six time points of seven, 14, 28, 64, 90 and 180 days. Baird-Parker agar (BP agar), Kenner Fecal (KF), Salmonella–Shigella agar (SS agar), Violet Red Bile Glucose Agar (VRBGA), and Sabouraud Dextrose agar (SD agar) media were used for enumeration of the six spiked microorganisms including Staphylococcus aureus, Enterococcus faecalis, Salmonella Enteritidis, Enterobacter species and Escherichia coli, Fusarium oxysporum and Aspergillus flavus, respectively. Direct antibacterial activity of the composites was also determined. De novo frequencies of microorganisms in five saffron samples were at acceptable levels with dominance of fungi species. Nanosilver embedded packages accelerated the reduction in live microbial numbers in saffron samples and the efficacy was the best in packages containing 4000 ppm nanosilver particles. Nanosilver packaging can significantly reduce microbial burden of saffron.     

Iodine binding property of a ternary complex consisting of starch,protein, and free fatty acids

A ternary complex consisting of amylose, whey protein, and free fatty acids (FFA) has been identified in our previous investigations, and its iodine binding properties were investigated. After reaction with iodine solution, an absorption peak (λ_max) at 620 nm was shown for pure amylose whereas the λ_max decreased to 510 nm when amylose was first complexed with FFA. Interestingly, a λ_max of 550 nm with an intermediate absorbance was observed for the ternary complex indicating its intermediate spectrophotometric property. Consistently, the amount of iodine bound by the ternary complex was between free amylose and typical amylose–FFA complex from potentiometric titration indicating the amylose–FFA complex within the ternary complex is less compact and more space is left for iodine binding. This in-between property of the ternary complex suggests it can be used as a molecular carrier to accommodate a forth component in addition to its functional lipids carrying capability in food product development.     

Iodine concentration of milk in a dose–response study with dairy cows and implications for consumer iodine intake

Most feed is poor in iodine and iodine supplementation of cow's diets must guarantee milk iodine concentrations for humans that contribute to prevention of the deficiency and minimize the risk of exceeding an upper limit of iodine intake. Five Holstein cows were fed four iodine doses (via Ca(ΙO₃)₂·6H₂O). In four sequential 14-d periods, doses of 0.2 (basal diet), 1.3, 5.1, and 10.1 mg iodine kg^?1 diet dry matter (DM) were administered. Samples of milk were collected during each period; blood was also sampled from each cow for each iodine dosage. In an 18-d depletion period, a non-supplemented diet was provided. Iodine was determined by inductively coupled plasma-mass spectrometry. The iodine content of milk and serum reflected the iodine dosages in feed significantly. The levels for the four doses tested in milk were 101±32, 343±109, 1215±222, and 2762±852 μg iodine kg^?1. The total amount of iodine in milk per day was 30–40% of ingested supplemental iodine. Omitting additional iodine resulted in a short-term reduction of serum and milk iodine following an exponential decay function. The iodine supplementation of 0.5–1.5 mg kg^?1 diet DM represents the requirement of the cow, resulting in 100–300 μg iodine L^?1 milk, which optimally contributes to human supply. The maximum dietary levels of former and present EU legislations (10 and 5 mg iodine kg^?1 cow feed) increase the risk of iodine excess in humans.     

Chemical composition,angiotensin I-converting enzyme (ACE) inhibitory,antioxidant and antimicrobial activities of the essential oil from south Tunisian Ajuga pseudoiva Rob. Lamiaceae

The essential oil of Ajuga pseudoiva, collected from Tunisia, was analyzed using gas chromatography–mass spectroscopy. Thirty-two compounds accounting for 95.76% of the total oil were identified. Sesquiterpenes were found to be the most abundant components of A. pseudoiva oil. And they were mainly represented by viridiflorol (30.17%), germacrene B (9.26%) (α, β and γ)-eudesmol (8.11%) and aromadendrene (7.45%). The essential oil of A. pseudoiva showed radical scavengers activity (IC₅₀ = 0.72 mg/mL) and displayed lipid peroxidation inhibitory activity (IC₅₀ = 0.6 mg/mL). A. pseudoiva essential oil was also found to exhibit a dose-dependent ACE inhibitory activity with an IC₅₀ value of 65.5 μg/mL. Moreover, the antimicrobial activity of the essential oil was tested against 17 species of microorganisms, and the results obtained showed significant antibacterial activity against the Gram-positive and Gram-negative bacteria, with inhibition zones and minimal inhibitory concentration values of 14–32 mm and 84–137 μg/mL and 6–21 mm and 105–336 μg/mL, respectively. Higher activity was also found against several fungal strains.     

Distribution and interspecific correlation of root biomass density in an arid Elaeagnus angustifolia–Achnatherum splendens community

Main objective of this study was to determine the interspecific relationships between two dominant species in terms of root distribution in a typical arid tree-herbage (Elaeagnus angustifolia–Achnatherum splendens) community at Xidatan, Pingluo County, Ningxia Autonomous Region, Northwest China. Eight concentric zones (namely, Z1–Z8) were set from the bases of E. angustifolia individuals to the open lands and five soil profiles were excavated in each zone. Each soil profile was divided into five layers at the depths of 0–10 cm, 10–30 cm, 30–60 cm, 60–100 cm and 100–150 cm. Roots were collected for each species, and soil water content (SWC) and soil bulk density (SBD) were measured for each layer. We found noteworthy roots layer separation in the sub-canopy zones (Z1–Z4). The soil layers with highest fine root biomass density (FRBD) of A. splendens was primarily in the 0–10 cm, which were significantly shallower than those of E. angustifolia; whereas in the inter-canopy zones (Z5–Z8), inconsistent separation, or even overlapping of highest-FRBD-layers emerged between the two dominant species. Correlation analyses showed that negative correlations of FRBD between the two species mainly occurred in those soil layers with relatively higher FRBD and lower SWC. In contrast, positive correlations corresponded with relatively lower FRBD and higher SWC.