Preparation of Chitosan Nanocompositeswith a Macroporous Structure by Unidirectional Freezing and Subsequent Freeze-Drying

Chitosan is the N-deacetylated derivative of chitin, a naturally abundant mucopolysaccharide that consists of 2-acetamido-2-deoxy-β-d-glucose through a β (1→4) linkage and is found in nature as the supporting material of crustaceans, insects, etc. Chitosan has been strongly recommended as a suitable functional material because of its excellent biocompatibility, biodegradability, non-toxicity, and adsorption properties. Boosting all these excellent properties to obtain unprecedented performances requires the core competences of materials chemists to design and develop novel processing strategies that ultimately allow tailoring the structure and/or the composition of the resulting chitosan-based materials. For instance, the preparation of macroporous materials is challenging in catalysis, biocatalysis and biomedicine, because the resulting materials will offer a desirable combination of high internal reactive surface area and straightforward molecular transport through broad “highways” leading to such a surface. Moreover, chitosan-based composites made of two or more distinct components will produce structural or functional properties not present in materials composed of one single component. Our group has been working lately on cryogenic processes based on the unidirectional freezing of water slurries and/or hydrogels, the subsequent freeze-drying of which produce macroporous materials with a well-patterned structure. We have applied this process to different gels and colloidal suspensions of inorganic, organic, and hybrid materials. In this review, we will describe the application of the process to chitosan solutions and gels typically containing a second component (e.g., metal and ceramic nanoparticles, or carbon nanotubes) for the formation of chitosan nanocomposites with a macroporous structure. We will also discuss the role played by this tailored composition and structure in the ultimate performance of these materials.     

Yield, water use efficiency and nitrogen uptake in potato: influence of drought stress

Summary A lysimeter experiment was performed to study the optimal allocation of limited water supply in potatoes. Irrigation regimes equal to 40, 60 and 80% of maximum evapotranspiration (ET) were evenly applied over the crop cycle. Other treatments involved withholding 80 mm of irrigation, based on ET, beginning at each of three designated growth stages (tuber initiation, early and late tuber growth). An irrigated control treatment, restoring the entire ET, was included for comparison. Continuous drought stress reduced photosynthesis as irrigation volumes were reduced. Plant biomass and tuber yield decreased almost proportionally to water consumption, so that WUE was roughly constant. N uptake was highest in the control and in 80% ET treatment. Withholding water during tuberisation severely hindered plant physiological processes and penalized tuber yield. Reductions in photosynthesis, total biomass and yield were the greatest when drought was imposed during tuber initiation. The earliest stress resulted in the lowest WUE and N uptake. A new crop water stress index (SI) was proposed, which combines atmospheric demand for water and canopy temperature.     

Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11

The infection of stored apples by the fungus Penicillium expansum causes the contamination of fruits and fruit-derived products with the mycotoxin patulin, which is a major issue in food safety. Fungal attack can be prevented by beneficial microorganisms, so-called biocontrol agents. Previous time-course thin layer chromatography analyses showed that the aerobic incubation of patulin with the biocontrol yeast Rhodosporidium kratochvilovae strain LS11 leads to the disappearance of the mycotoxin spot and the parallel emergence of two new spots, one of which disappears over time. In this work, we analyzed the biodegradation of patulin effected by LS11 through HPLC. The more stable of the two compounds was purified and characterized by nuclear magnetic resonance as desoxypatulinic acid, whose formation was also quantitated in patulin degradation experiments. After R. kratochvilovae LS11 had been incubated in the presence of (13)C-labeled patulin, label was traced to desoxypatulinic acid, thus proving that this compound derives from the metabolization of patulin by the yeast. Desoxypatulinic acid was much less toxic than patulin to human lymphocytes and, in contrast to patulin, did not react in vitro with the thiol-bearing tripeptide glutathione. The lower toxicity of desoxypatulinic acid is proposed to be a consequence of the hydrolysis of the lactone ring and the loss of functional groups that react with thiol groups. The formation of desoxypatulinic acid from patulin represents a novel biodegradation pathway that is also a detoxification process.     

Surgical technique, postoperative complications and outcome in 14 dogs treated for hydrocephalus by ventriculoperitoneal shunting

Objective: To report frequency and type of complications, and outcome in dogs with severe neurologic signs secondary to internal, suspected obstructive hydrocephalus treated by ventriculoperitoneal (VP) shunting. Study Design: Case series. Animals: Dogs (n=14). Methods: Medical records (2001–2006) was reviewed for dogs that had VP shunting. Inclusion criteria were complete medical record, progressive forebrain signs unresponsive to medical treatment, normal metabolic profile, negative antibody titers and/or cerebrospinal PCR for Toxoplasma gondii, Neospora caninum, and canine distemper virus, magnetic resonance images of the brain, confirmed diagnosis of VP shunting, and follow‐up information. Results: Hydrocephalus was idiopathic in 5 dogs and acquired (interventricular tumors, intraventricular hemorrhage, inflammatory disease) in 9 dogs. Four dogs developed complications 1 week to 18 months postoperatively, including ventricular catheter migration, infection, shunt under‐drainage, kinking of the peritoneal catheter, valve fracture, and abdominal skin necrosis. Three of these dogs had 1 or more successful revision surgeries and 1 dog was successfully treated with antibiotics. All, but 1 dog, were discharged within 1 week of surgery, and had substantial neurologic improvement. Median survival time for all dogs was 320 days (1–2340 days), for dogs with idiopathic hydrocephalus, 274 (60–420) days and for dogs with secondary hydrocephalus, 365 (1–2340) days. Conclusions: VP shunting was successful in relieving neurologic signs in most dogs and postoperative complications occurred in 29%, but were resolved medically or surgically.     

Effects of arbuscular mycorrhizal inoculation on growth,yield, nitrogen,and phosphorus nutrition of nodulating bean varieties in two soil substrates of contrasting fertility

A pot experiment was conducted to study the response of nodulated bean (Phaseolus vulgaris L.), vars. Ceca and Montalbán, to inoculation with the arbuscular‐mycorrhizae (AM) Glomus spurcum strain IES‐3 and G. mosseae strain IES‐8 in two non‐sterilized non‐amended soil substrates (S‐l and S‐2) of contrasting fertility. S‐l was an acid soil (pH 5.0) with 77% exchangeable aluminum (Al), low available phosphorus (P) (3.5 ug g^‐1 dry soil) and 1.8% organic matter. S‐2 was a soil with pH 7.0, 7.9 μg P g^‐1 dry soil, 9.6% organic matter and no exchangeable Al. Plants growing in S‐1 (S‐1 plants) and S‐2 (S‐2 plants) soils were sampled at the beginning of the flowering stage. S‐1 and S‐2 plants were nodulated by the rhizobial populations native of each soil. S‐1 plants of both varieties did not respond to AM inoculation in term of shoot and root mass, P content, relative abundance of ureides (RAU) and seed yields. Only the total chlorophyll, chlorophyll‐a, chlrophyll‐a/b ratio and nodule mass were significantly enhanced in AM‐inoculated plants. Increased total reducing sugars concentration was detected in roots of S‐1 plants inoculated with G. mosseae. S‐2 plants displayed significantly higher shoot and nodule mass as well as increased total chlorophyll, chlorophyll‐a, chlorophyll‐a/b ratio and P content than S‐1 plants, regardless of the variety and AM inoculation. S‐2 plants of the var Ceca showed a two fold increase in seed yields but similar RAU values (>60%) when compared to S‐1 plants of the same variety. In contrast, the significant increase in RAU detected in S‐2 plants of the var Montalban was not translated into higher seed yields. In S‐2, the productivity of plants of the var Ceca doubled that of the var Montalban. For both bean varieties the highest significant P content and seed yield were observed exclusively in S‐2 plants inoculated with G. mosseae. This Glomelean strain enhanced the sink‐source ratio of the S‐2 plants as evidenced by the higher total reducing sugar concentration in the root mass. Arbuscular‐mycorrhizae inoculation significantly decreased the acid phosphatase activity in the rhizosphere of S‐1 and S‐2 plants, respectively, pointing toward a negative effect of foreign AM on the native microbial biomass. The effectiveness of the rhizobial populations native of each soil type and the weak response elicited by G. mosseae in S‐2 plants do not justify, at present, the inclusion of foreign inocula in the bean crops carried out at S‐1 and S‐2 soils of the Sucre State of Venezuela. Results also indicated the higher adaptability of var Ceca to conditions prevailing in S‐1 and S‐2.     

Effect of Copper on Antioxidant Enzyme Activities and Mineral Nutrition of White Lupin Plants Grown in Nutrient Solution

We analyzed the effect of different copper (Cu) concentrations (0.10, 0.15, 0.20 and 0.35 mM) and time (1 day to 9 days) on several growth and biochemical parameters of roots and shoots of white lupin plants (Lupinus albus cv Estoril) grown in nutrient solution. A significant decrease in leaf fresh weight and leaf area was detected. Copper accumulated in the roots, and an impairment of nutrient translocation was only observed after six days at the highest Cu concentrations applied. A transient increase in the activity of polyphenoloxidase (EC 1.10.3.1) enforces a role for lignification as a defense strategy under enhanced Cu levels. The activities of several antioxidative enzymes were enhanced after Cu application. Our results indicate that Lupinus albus cv ‘Estoril’ is a rather resistant plant that can cope with moderate concentrations of copper, mostly by controlling up to a certain point, the uptake of excessive amounts of this metal.     

Fluorine immission to acid soil in the vicinity of an aluminium smelter in Galicia (NW Spain) and its influence on aluminium dynamics

Purpose

The aluminium smelter located in the northern coast of Galicia (NW Spain) is a source of fluoride pollution. Previous papers Gago et al. (Commun Soil Sci Plant Anal 32:2503–2517, 2001, Fluoride 35:110–121, 2002) have identified an area of 3-km radius around the factory being affected by fluoride emissions. This study aims to investigate the influence of fluoride emissions on fluorine concentration and speciation as well as on aluminium speciation in soils in the vicinity of the smelter.

Materials and methods

PVC cylinders containing soil material from the A horizon of an unpolluted forest soil, similar to that in the vicinity of the factory, were placed within the A horizon of soils surrounding the smelter. The cylinders were placed at various distances (up to 3 km) from the factory in the direction of predominant winds. The PVC columns were removed after 6 months, and the soil material inside was divided into two slices (0–10 and 10–20 cm) and analysed for pH, total and available fluorine, exchangeable cations, effective cation exchange capacity (CEC) and oxalate-extracted aluminium. The soil solution was mimicked by aqueous extracts and analysed for pH, electrical conductivity, Ca, Mg, Na, K, total and free fluorine, total Al, acid-soluble Al, non-labile Al, labile Al and labile Al species.

Results and discussion

In the area closest to the factory (0.5 km), total and available fluorine in the upmost 10 cm soil increased by twofold and eightfold, respectively, while soluble fluoride increased by fourfold. Increases were also observed in exchangeable Ca, Mg, Al and CEC. Noncrystalline Al increased significantly at any distance from the smelter. Fluorine, pH, sulphate, aluminium and organic matter increased in the soil solution of surface (<10 cm) samples closest to the factory. Monomeric Al-organic matter complexes prevailed in the liquid phase at 0.5–1 km from the factory whereas inorganic Al complexes did at distances over 1 km. Among the inorganic Al complexes, fluoride complexes prevailed at distances less than 1 km and hydroxylated complexes at distances greater than 1 km. Al³⁺ concentration was always very low.

Conclusions

Emissions from an aluminium smelter led to increased concentrations of fluorine and various forms of aluminium in the nearby soil. These emissions mobilised aluminium, organic matter and probably sulphate from the soil. In the closest neighbourhood of the smelter, it was found that most soluble aluminium was bound to organic matter, hence having low toxicity, while at distances farther than 1 km Al-F and Al-OH complexes prevailed.     

Two outer membrane proteins are bovine lactoferrin-binding proteins in Mannheimia haemolytica A1

Mannheimia haemolytica is a Gram negative bacterium that is part of the bovine respiratory disease, which causes important economic losses in the livestock industry. In the present work, the interaction between M. haemolytica A1 and bovine lactoferrin (BLf) was studied. This iron-chelating glycoprotein is part of the mammalian innate-immune system and is present in milk and mucosal secretions; Lf is also contained in neutrophils secondary granules, which release this glycoprotein at infection sites. It was evidenced that M. haemolytica was not able to use iron-charged BLf (BholoLf) as a sole iron source; nevertheless, iron-lacked BLf (BapoLf) showed a bactericidal effect against M. haemolytica with MIC of 4.88 ± 1.88 and 7.31 ± 1.62 μM for M. haemolytica strain F (field isolate) and M. haemolytica strain R (reference strain), respectively. Through overlay assays and 2-D electrophoresis, two OMP of 32.9 and 34.2 kDa with estimated IP of 8.18 and 9.35, respectively, were observed to bind both BapoLf and BholoLf; these OMP were identified by Maldi-Tof as OmpA (heat-modifiable OMP) and a membrane protein (porin). These M. haemolytica BLf binding proteins could be interacting in vivo with both forms of BLf depending on the iron state of the bovine.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-016-0378-1) contains supplementary material, which is available to authorized users.     

Lipid droplet distribution of immature canine oocytes in relation to their size and the reproductive stage

This study investigated the distribution of lipid droplets (LD) in immature canine oocytes in relation to their size and the reproductive stage. Oocytes were collected from the ovaries of bitches at different estrous stages, divided according to their size (110‐120 µm; >120 µm), and stained with Nile Red to detect lipid droplet distribution. At the follicular phase most of the oocytes displayed a diffuse pattern of LD distribution, whereas at anestrus and luteal phase oocytes showed LD mainly in a peripheral/ perinuclear LD distribution. A significantly higher intensity of LD has been recorded in the oocytes > 120 µm compared to those of smaller size (110 ‐ 120 µm) at all stages of the estrous cycle. At follicular phase, oocytes > 120 µm displayed LD intensity similar to that of oocytes > 120 µm at luteal phase and higher compared to the oocytes of the other groups.