Effect of feeding wheat middlings and calcium lignosulfonate as pellet binders on pellet quality growth performance and lipid peroxidation in broiler chickens.

Basal diets supplemented with 4 kg Ca-LS/ton of diet. Pellet quality characteristics (per cent fines, the present study was conducted to evaluate the influence of wheat middlings (WM) and calcium lignosulfonate (Ca-LS) as pellet binders on the pellet quality characteristics, growth performance, blood parameters, nutrients digestibilities, lipid peroxidation and muscle fatty acids profile in Egyptian broiler strain. A total of 3,120 broiler chicks at 1-day of age were divided randomly into three experimental treatments with eight replicates (130 each). The first treatment was fed the basal pelleted diets without any additives, the second treatment was fed diets including 50 kg WM/ton of diet and the third treatment was fed per cent pellets, and pellet durability index) were significantly improved in WM and Ca-LS treatments compared with the control. Body weight gain was significantly increased, while feed intake was significantly decreased resulting in improving of feed conversion ratio significantly in WM group in comparison with control and Ca-LS groups (p < .05). Nutrients apparent digestibility (dry matter, crude protein and crude fibre) were significantly improved by inclusion of WM compared with control and Ca-LS. Plasma total cholesterol, and uric acid concentrations were significantly decreased by dietary WM in comparison with control and Ca-LS experimental groups. Furthermore, linoleic, alpha-linolenic and arachidonic acids contents in breast muscle were significantly increased by WM and Ca-LS, while, muscle malondialdehyde concentration was significantly decreased. It could be concluded that inclusion of WM and Ca-LS can improve pellet quality characteristics, and WM (at a level of 50 kg/ton) had positive effects on growth performance, nutrients digestibilities, lipid peroxidation and fatty acids profile in Egyptian broiler strain.

The effects of lignocellulose supplementation on laying performance, egg quality parameters, aerobic bacterial load of eggshell, serum biochemical parameters, and jejunal histomorphological traits of laying hens.

This study was performed to investigate the effects of lignocellulose supplementation (LS) on performance parameters, egg quality, aerobic bacterial load of eggshell, serum biochemical parameters, and jejunal histomorphological traits of laying hens between 18 and 38 wk of age. A total of 640 pullets at 16 wk of age were allotted to 4 treatment groups as 0 kg (control, CONT), 0.5 kg, 1 kg, and 2 kg LS per ton of feed. Body weight (BW), daily feed intake, egg production (EP), egg weight (EW), and efficiency of feed utilization (EF) were determined as the mean of each 3-wk period between 18 and 38 wk of age. Laying hens in the 1 kg LS group had a higher BW mean (1632.1 g, P < 0.001). The highest mean value of EP and EW were observed in 1 kg LS group (81.8% and 57.3 g, respectively), whereas the lowest values were found in the 2 kg LS group (78.6% and 54.4 g, respectively, P < 0.001). The mean of EF was the lowest in the 1 kg LS group (2.72, P < 0.001). There was a decline in eggshell breaking strength and eggshell thickness in the 2 kg LS, when compared with the 0.5 and 1 kg LS groups (P < 0.001). The total aerobic bacterial load of the eggshell was the lowest in the 1 kg LS group (4.7 log10 cfu/mL). The level of aspartate amino transferase and alanine amino transferase showed an increment in both the CONT and 2 kg LS groups (P < 0.001). The high level of LS (2 kg per ton of feed) caused a decline in the levels of IgY, IgA and IgM, when compared to the 0.5 and 1 kg LS groups (P < 0.001). Laying hens in 0.5 and 1 kg LS groups had longer villus height (1335.9 µm) in the jejunum than the others (P < 0.001). These findings showed that the 1 kg LS per ton of feed improved EP and EW, eggshell quality, immunoglobulin levels and intestinal morphology, and decreased the total aerobic bacterial load.

Lactoferrin/Hyaluronic acid double-coated lignosulfonate nanoparticles of quinacrine as a controlled release biodegradable nanomedicine targeting pancreatic cancer.

Quinacrine is an antimalarial drug that was repositioned for treatment of cancer. This is the first work to enhance quinacrine activity and minimize its associated hepatotoxicity via loading into bio-degradable, bio-renewable lignosulfonate nanoparticles. Particles were appraised for treatment of pancreatic cancer, one of the most life-threatening tumors with a five-year survival estimate. Optimum nanocomposites prepared by polyelectrolyte interaction exhibited a particle size of 138 nm, a negative surface charge (-28 mV) and a pH dependent release of the drug in an acidic environment. Ligands used for active targeting (lactoferrin and hyaluronic acid) were added to nanoparticles' surface via layer by layer coating technique. The highest anticancer activity on PANC-1 cells was demonstrated with dual active targeted particles (3-fold decrease in IC50) along with an increased ability to inhibit migration and invasion of pancreatic cancer cells. In vivo studies revealed that elaborated nanoparticles particles showed the highest tumor volume reduction with enhanced survival without any toxicity on major organs. In conclusion, the elaborated nanoparticles could be considered as a promising targeted nanotherapy for treatment of pancreatic cancer with higher efficacy& survival rate and lower organ toxicity.

Effects of soybean hulls and lignocellulose on performance, nutrient digestibility, microbial metabolites and immune response in piglets.

A feeding trial with 96 piglets was performed to investigate the effect of added soluble (SDF) and insoluble dietary fibre (IDF) sources on performance, apparent total tract digestibility (ATTD), concentration of microbial metabolites and pro-inflammatory marker genes as indicators for immune response. Piglets were allotted to four treatments (T): T1 control, T2 with soybean hulls (IDF/SDF: 8.35) and T3 and T4 with two different kinds of lignocellulose (IDF/SDF: >70). Diets were isofibrous for their value of total dietary fibre to underline the particular physicochemical properties of fibre sources. No differences were observed regarding average daily feed intake, average daily gain (ADG), feed conversion ratio and body weight, while T2 expressed higher ADG in the grower phase (day 14-54) vs. T3. Soybean hulls (T2) resulted in higher ATTD of dry matter and organic matter vs. T4; ether extract vs. T1 and neutral detergent fibre vs. T1, T2 and T3. The concentration of short chain fatty acids did not differ among treatments. Ileal digesta in T2 generated higher amounts of cadaverine vs. T3 and T4, likewise T1 vs. T4. Finally, no impact on immune response was detected. In conclusion, soybean hulls affected ATTD positively and lignocellulose prevented the formation of cadaverine, no overall direct response of SDF nor of IDF for the inclusion level  were observed.

Multifunctional Bioplastics Inspired by Wood Composition: Effect of Hydrolyzed Lignin Addition to Xylan-Cellulose Matrices.

Multifunctional bioplastics have been prepared by amorphous reassembly of cellulose, hemicelluloses (xylan), and hydrolyzed lignin. For this, the biopolymers were dissolved in a trifluoroacetic acid-trifluoroacetic anhydride mixture and blended in different percentages, simulating those found in natural woods. Free-standing and flexible films were obtained after the complete evaporation of the solvents. By varying xylan and hydrolyzed lignin contents, the physical properties were easily tuned. In particular, higher proportions of hydrolyzed lignin improved hydrodynamics, oxygen barrier, grease resistance, antioxidant, and antibacterial properties, whereas a higher xylan content was related to more ductile mechanical behavior, comparable to synthetic and bio-based polymers commonly used for packaging applications. In addition, these bioplastics showed high biodegradation rates in seawater. Such new polymeric materials are presented as alternatives to common man-made petroleum-based plastics used for food packaging.

Growth performance, pH value of gizzard, hepatic enzyme activity, immunologic indicators, intestinal histomorphology, and cecal microflora of broilers fed diets supplemented with processed lignocellulose.

This study was performed to investigate the hypothesis that supplementation of processed lignocellulose (PL) in the diets of broilers has a positive effect on growing performance, pH value of gizzard, hepatic enzyme activity, immunologic indicators, histomorphological character of small intestine, and cecal microflora populations. A total of 720 one-day-old Ross 308 broiler chicks were allotted to 4 treatment groups and fed maize-soybean meal based diets. The basal diet was supplemented with PL with an amount of 0 kg (control), 0.5 kg, 1 kg, and 2 kg per ton feed. Growing performance parameters, were determined weekly until 35 D of age. Blood samples for enzyme activities and immunoglobulins, jejunum and cecum samples for histomorphological characters for villus growth, and microbial population were collected from 12 broilers from each group. At 35 D of age, body weight of broilers supplemented with 1 kg of PL was found to be the highest with a value of 2305.0 g, when compared to the broilers supplemented with control, 0,5 and 2 kg of PL groups (2154.0, 2201.0, and 2141.7 g, respectively, P = 0.001). An increased activity of aspartate amino transferase (AST) was observed in the control and 1 kg PL supplementation groups (633.6 and 597.4 IU/L, respectively), whereas alkaline phosphatase (ALP) activity was the highest in the control group (5404 IU/L, P < 0.05). Broilers in the control group had the lowest level of IgY and IgA (122.2 and 25.8 mg/dL, respectively, P < 0.05). Villus height increased by 22.0%, 40.7%, and 34.8% in 0.5, 1, and 2 kg PL supplementation groups, respectively, when compared to the control (P < 0.001). The processed lignocellulose supplemented as 1 kg of PL decreased the average count of Staphylococcaceae, E. coli, and Enterobacteriaceae, whereas it increased the population of Lactobacillus spp. in the cecum (P < 0.05). These data indicate that the supplementation of processed lignocellulose had positive effects for performance via changes in hepatic enzyme activities, immunoglobulin levels, villus growth in jejunum, and microflora in cecum.

Effects of reducing growth rate via diet dilution on bone mineralization, performance and carcass yield of coccidia-infected broilers.

Coccidiosis and rapid growth rate (GR) compromise bone mineralization in modern broilers. We tested the hypothesis that reducing GR via diet dilution during peak bone development will improve bone mineralization in both infected and uninfected broilers. A total of 384 male Ross 308 chicks were allocated to a basal grower diet (3,107 kcal/kg ME and 19.4% CP) diluted with 0, 5, 10, or 15% lignocellulose (n = 12 pens/treatment, 8 birds/pen) at day 10 of age. Prior to this, birds in each group received half the intended diet-dilution levels (day 8 to 10 of age) and a common starter diet (day 1 to 7 of age). At day 13 of age (day 0 post-infection, pi), birds were orally inoculated with either 7,000 sporulated Eimeria maxima oocysts (I) or water (C), forming a 4 diet-dilution level × 2 infection status factorial experiment. Performance was measured over 12 days pi and scaled to BW at infection (day 0 pi) to account for a priori BW differences. At day 12 pi (day 25 of age), 1 bird/pen (a total of 6 birds/treatment) was sampled to assess tibia and femur mineralization relative to BW, and carcass yield. There was no interaction (P > 0.05) between infection status and diet-dilution level on ADFI/BW measured over day 1 to 12 pi, or on any bone variable. ADG/BW pi decreased (P < 0.01) with diet dilution amongst C birds, but was statistically similar (P > 0.05) amongst I birds. I compared to C birds had reduced breast meat (P < 0.05) and eviscerated carcass yield (P < 0.01), femur (P < 0.05) and tibia (P < 0.01) breaking strength (BS), and femur ash weight (AW) (P < 0.05). Diet dilution did not affect carcass yield, but improved femur BS (P < 0.001), and tended to improve (P < 0.1) femur and tibia AW. Overall, diet dilution significantly affected femur, more than tibia, variables: relative BS, robusticity index, and ash percentage. Reducing GR affected broiler long bone mineralization to a similar degree in the presence or absence of coccidiosis.

Effect of calcium lignosulfonate supplementation on metabolic profiles of confined lambs.

This study aimed to evaluate the effect of calcium lignosulfonate associated with whole cottonseed in high-concentrate diets for sheep. Eight Dorper crossbred sheep with an average live weight of 42.5 ± 1.70 kg were assigned to two 4 × 4 Latin squares. The following experimental diets were evaluated: control diet (without calcium lignosulfonate) and diets with inclusion of 50, 100, and 150 g of calcium lignosulfonate/kg fresh matter. Diets were composed of soybean meal, ground corn, and whole cottonseed. Feed intake, digestibility, metabolic characteristics, and feeding behavior were evaluated. The intake of nutritional components did not show significant differences as a function of the lignosulfonate levels in the diet; however, the increase in calcium lignosulfonate levels linearly decreased the dry matter digestibility. Rumen ammonia nitrogen concentrations decreased linearly as the lignosulfonate levels in the diets were increased. There was no effect of lignosulfonate levels on blood parameters or feeding behavior of the animals. The use of lignosulfonate associated with cottonseed decreases the digestibility of dry matter and the concentration of rumen ammonia nitrogen, but does not change the intake of nutritional components, the blood parameters, or the feeding behavior of sheep.

A novel lignin-based nanofibrous dressing containing arginine for wound-healing applications.

Nanofiber-based wound dressings have attracted much attention in wound care owing to their unique properties such as high aspect ratio and three-dimensional structure. Arginine is a precursor of nitric oxide that plays an important role in the wound-healing process. Therefore, in this study, we have developed a gel which contains lignin nanofibers (Lig-NFs) that were surface modified by arginine molecules via electrostatic interaction (Arg-Lig-NF gel). The effect of pH on the amount of arginine attached on Lig-NF surface was evaluated at three different pH values-5, 6, and 7. Fourier transform infrared spectroscopy and zeta potential of Lig-NFs before and after surface modification confirmed the surface modification of Lig-NFs with arginine molecules. The optimum gel composed of uniform Arg-Lig-NFs with diameter ranging from 100 to 250 nm. There was 184.60 ± 4.85 mg arginine in each gram of optimum gel. The release of arginine from Arg-Lig-NF gel showed a sustained release manner, and about 86.28 ± 3.50% of attached arginine were released after 24 h. Moreover, the optimum gel presented suitable viscosity and spreadability for topical application. The in vivo full thickness wound-healing assay carried out in rats demonstrated that the optimum Arg-Lig-NF gel can accelerate wound closure and increase re-epithelialization, collagen deposition, and angiogenesis significantly in Arg-Lig-NF gel-treated wounds compared to Lig-NF gel and arginine solution. Overall, these findings demonstrate that Arg-Lig-NF gel can be a promising material for the future development of effective hydrocolloid wound dressings used in the treatment of acute and chronic wounds.

Coarse particle inclusion and lignocellulose-rich fiber addition in feed benefit performance and health of broiler chickens.

Measures to improve gut health and nutrient digestibility have been sought due to in-feed antibiotics being phased out in poultry. The appropriate physical structure of feed ingredients and addition of dietary fiber may be beneficial in enhancing gut health in poultry. In this study, the effect of a lignocellulose-rich fiber source and corn particle size on broiler performance, gizzard development, nutrient digestibility, cecal microflora, and litter quality was evaluated. A total of 684 day-old male Ross 308 chicks were randomly allocated to 6 treatments with 6 replicate pens, each housing 19 birds. A 2 × 3 factorial arrangement of treatments was applied with the factors of: corn particle size (coarse: 2,982 µm or fine: 941 µm geometric mean diameter), and 3 levels of lignocellulose (0%, 1% or 2%). Significant particle size × fiber interaction was observed for feed conversion ratio (FCR) at d 10 (P < 0.05). The birds fed coarsely ground corn (CGC) had lower FCR than those fed finely ground corn (FGC) only at 2% of lignocellulose but not at 1% or no lignocellulose addition. Birds fed FGC were heavier (P < 0.001) at d 10. In contrast, at d 24 and 35, birds fed CGC had lower FCR than those fed FGC (P < 0.05). Ileal gross energy and protein digestibility increased in birds fed CGC compared with those fed FGC at d 24 (P < 0.05). Relative gizzard weight was higher (P < 0.05) on d 24 and 35 in birds fed CGC as compared to those fed FGC. Birds consumed 2% dietary lignocellulose had decreased counts of cecal Clostridium spp. compared to those with 1% lignocellulose (P < 0.05) at d 24. On d 35, both levels of lignocellulose had significantly decreased (P < 0.05) litter moisture content compared to the control. In conclusion, birds fed pelleted diets containing CGC exhibited improved FCR, and increased nutrient digestibility, which may have been caused by larger gizzards. Furthermore, dietary lignocellulose addition is beneficial to litter quality.

Supplementing the feeds of layer pullets, at different ages with two different fiber sources improves immune function.

Two experiments were conducted to study the effects of lignocellulose supplementation on immune function in layer pullets at different stages of growth. Four-wk-old pullets (Experiment 1) were fed a control, diet (Diet C); Diet C plus 1% mixed soluble/insoluble fiber (Diet MF), or plus 1% insoluble fiber (Diet IF). At 7.5 wk-of-age, heterophil phagocytosis, and oxidative burst in Groups MF (328.5 beads/100 cells; 4,330.0 ΔRFU; relative fluorescent units) and IF (350.3; 5,264.4) were greater (P < 0.05) than Controls (303.4; 3,509.0). At 8 wk-of-age, Group MF and IF relative weights of bursa of Fabricius (0.57 g/100 g BW; 0.58 g /100 g BW), thymus glands (0.77; 0.78), and areas of Peyer's patches (PP) (2.7 cm2; 2.9 cm2) were higher (P < 0.05) than Controls (bursa, 0.50 g; thymus, 0.70 g; PP area, 1.8 cm2). In Experiment 2, 10-wk-old pullets were fed a control diet or diets containing 1.5% MF or IF for 8 wk. At 14 wk-of-age IF pullets had higher (P < 0.05) heterophil phagocytosis efficiency (447.9 beads/100 cells) than Controls (376.4) and MF and IF had greater (P < 0.05) oxidative burst (1,302.9 and 1,857.7 ΔRFU) than Controls (744.1). At 17 wk-of-age MF and IF had increased (P < 0.05) proliferation of T-lymphocytes (ConcanavalinA-stimulated) (100.4 and 103.1% of unstimulated cells) and B-lymphocytes (lipopolysaccharide-stimulated) (122.4 and 129.0) than Controls (ConA, 79.4; lipopolysaccharide, 106.6). At 18 wk-of-age, IF pullets were heavier (1,607.5 g, P < 0.05) than Controls (1,506.5 g), had heavier (P < 0.05) bursa of Fabricius (1.12 g) than MF and Control groups (0.98 g; 0.92 g) and cecal tonsils of MF (0.38 g) and IF (0.39 g) weighted more (P < 0.05) than Controls (0.33 g). Number of jejunal and ileal PP (10.0) in IF pullets was higher (P < 0.05) than Controls (7.1). These results indicate that both MF and IF can improve development of the immune system of young and grower pullets during periods of maturation and involution of lymphoid organs.

Lignin-graft-Polyoxazoline Conjugated Triazole a Novel Anti-Infective Ointment to Control Persistent Inflammation.

Lignin, one of the most abundant renewable feedstock, is used to develop a biocompatible hydrogel as anti-infective ointment. A hydrophilic polyoxazoline chain is grafted through ring opening polymerization, possess homogeneous spherical nanoparticles of 10-15 nm. The copolymer was covalently modified with triazole moiety to fortify the antimicrobial and antibiofilm activities. The hydrogel was capable of down regulating the expression level of IL-1ß in LPS induced macrophage cells, and to cause significant reduction of iNOS production. It supported cellular anti-inflammatory activity which was confirmed with luciferase assay, western blot, and NF-κB analysis. This novel lignin-based hydrogel tested in-vivo has shown the abilities to prevent infection of burn wound, aid healing, and an anti-inflammatory dressing material. The hydrogel reported here provides a new material platform to introduce a cost-effective and efficient ointment option after undertaking further work to look at its use in the area of clinical practice.

Model-based characterisation of growth performance and l-lactic acid production with high optical purity by thermophilic Bacillus coagulans in a lignin-supplemented mixed substrate medium.

Three Bacillus coagulans strains were characterised in terms of their ability to grow in lignin-containing fermentation media and to consume the lignocellulose-related sugars glucose, xylose, and arabinose. An optical-density high-throughput screening was used for precharacterisation by means of different mathematical models for comparison (Logistic, Gompertz, Baranyi, Richards & Stannard, and Schnute). The growth response was characterised by the maximum growth rate and lag time. For a comparison of the screening and fermentation results, an unstructured mathematical model was proposed to characterise the lactate production, bacterial growth and substrate consumption. The growth model was then applied to fermentation procedures using wheat straw hydrolysates. The results indicated that the unstructured growth model can be used to evaluate lactate producing fermentation. Under the experimental fermentation conditions, one strain showed the ability to tolerate a high lignin concentration (2.5g/L) but lacked the capacity for sufficient pentose uptake. The lactate yield of the strains that were able to consume all sugar fractions of glucose, xylose and arabinose was ∼83.4%. A photometric measurement at 280nm revealed a dynamic change in alkali-lignin concentrations during lactate producing fermentation. A test of decolourisation of vanillin, ferulic acid, and alkali-lignin samples also showed the decolourisation performance of the B. coagulans strains under study.

In vitro evaluation of biodegradable lignin-based nanoparticles for drug delivery and enhanced antiproliferation effect in cancer cells.

Currently, nanosystems have been developed and applied as promising vehicles for different biomedical applications. We have developed three lignin nanoparticles (LNPs): pure lignin nanoparticles (pLNPs), iron(III)-complexed lignin nanoparticles (Fe-LNPs), and Fe3O4-infused lignin nanoparticles (Fe3O4-LNPs) with round shape, narrow size distribution, reduced polydispersity and good stability at pH 7.4. The LNPs showed low cytotoxicity in all the tested cell lines and hemolytic rates below 12% after 12 h of incubation. Additionally, they induced hydrogen peroxide production in a small extent and time-dependent manner, and the interaction with the cells increased over time, exhibiting a dose-dependent cell uptake. Concerning the drug loading, pLNPs showed the capacity to efficiently load poorly water-soluble drugs and other cytotoxic agents, e.g. sorafenib and benzazulene (BZL), and improve their release profiles at pH 5.5 and 7.4 in a sustained manner. Furthermore, the BZL-pLNPs presented an enhanced antiproliferation effect in different cells compared to the pure BZL and showed a maximal inhibitory concentration ranging from 0.64 to 12.4 µM after 24 h incubation. Overall, LNPs are promising candidates for drug delivery applications, and the superparamagnetic behavior of Fe3O4-LNPs makes them promising for cancer therapy and diagnosis, such as magnetic targeting and magnetic resonance imaging.

Uso de indicadores indigestíveis obtidos in situ e in vivo para determinar a digestibilidade de nutrientes em equinos / The use of indigestible markers obtained in situ and in vivo to determine digestibility of nutrients in equine

Objetivou-se estudar a viabilização dos indicadores internos, celulose (CELi) e lignina (LIGi) indigestíveis, obtidos pelas técnicas in situ e in vivo, para predizer a digestibilidade de nutrientes em equinos. Foram utilizados quatro cavalos castrados, por meio de delineamento em blocos ao acaso. Os tratamentos consistiram de metodologias de avaliação de digestibilidade, como a coleta total de fezes (CT) e o uso de indicadores internos, CELi e LIGi, obtidos pelas técnicas in situ (IS) na cavidade ruminal de bovinos e in vivo (IV) nos equinos, por meio da técnica do saco de náilon móvel (TSNM), totalizando cinco tratamentos. A ingestão diária de MS foi de 2,0 por cento do peso vivo de feno de coast-cross. A CELi-IV promoveu a melhor taxa de recuperação do indicador, igualando-se à CT, em 100,31 por cento, enquanto a LIGi resultou nas piores taxas, na média 76,45 por cento, para ambas as técnicas (P<0,05). As estimativas dos coeficientes de digestibilidade dos nutrientes foram adequadamente preditas pela CELi, obtidos IS e IV em que os valores observados foram de 46,41, 48,16, 46,90, 47,92 e 45,51 por cento para MS, MO, PB, FDN e EB, respectivamente. Concluiu-se que a CELi pode ser obtida in vivo por meio da TSNM em equinos, para predizer a digestibilidade de nutrientes...

The objective of this paper was to study the use of the internal indigestible cellulose (CELi) and lignin (LIGi) markers, obtained in situ or in vivo, to predict the apparent digestibility of nutrients in horses. Four castrated horses were utilized in a randomized blocks experimental design. Treatments consisted of different digestibility determination methodologies: total feces collection (TC) and use of CELi and LIGi internal markers obtained by the in situ (IS) incubation in bovine rumen and in vivo (IV) by the mobile nylon bag technique (MNBT) with horses, resulting in five treatments. The diet consisted exclusively of coast-cross hay and the average daily dry matter intake corresponded to 2.0 percent of the BW of the horses. The CELi-IV resulted in the best marker recuperation rate (100.31 percent), being similar to the control method; while the LIGi resulted in low recuperation rates (76.45 percent) for both techniques (P<0.05). The nutrients' digestibility coefficients were effectively predicted by CELi, obtained in situ or in vivo, with average values of 46.41, 48.16, 46.90, 47.92 and 45.51 percent for DM, organic matter, crude protein, neutral detergent fiber and crude energy, respectively. It can be concluded that CELi may be obtained in vivo by MNBT in horses to predict the nutrients' digestibility coefficients...

Ultrasound driven assembly of lignin into microcapsules for storage and delivery of hydrophobic molecules.

Oil-filled microcapsules of kraft lignin were synthesized by first creating an oil in water emulsion followed by a high-intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. The rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. As further evidence in support of our rationale, confocal and optical microscopies demonstrated the uniformly spherical morphology of the created lignin microparticles. The detailed elucidation of the cross-linking processes was carried out using gel permeation chromatography (GPC) and quantitative (31)P NMR analyses. The ability of lignin microcapsules to incorporate and release Coumarin-6 was evaluated in detail. In vitro studies and confocal laser scanning microscopy analysis were carried out to assess the internalization of capsules into Chinese hamster ovary (CHO) cells. This part of our work demonstrated that the lignin microcapsules are not cytotoxic and readily incorporated in the CHO cells.

Lignin nanotubes as vehicles for gene delivery into human cells.

Lignin nanotubes (LNTs) synthesized from the aromatic plant cell wall polymer lignin in a sacrificial alumina membrane template have as useful features their flexibility, ease of functionalization due to the availability of many functional groups, label-free detection by autofluorescence, and customizable optical properties. In this report we show that the physicochemical properties of LNTs can be varied over a wide range to match requirements for specific applications by using lignin with different subunit composition, a function of plant species and genotype, and by choosing the lignin isolation method (thioglycolic acid, phosphoric acid, sulfuric acid (Klason), sodium hydroxide lignin), which influences the size and reactivity of the lignin fragments. Cytotoxicity studies with human HeLa cells showed that concentrations of up to 90 mg/mL are tolerated, which is a 10-fold higher concentration than observed for single- or multiwalled carbon nanotubes (CNTs). Confocal microscopy imaging revealed that all LNT formulations enter HeLa cells without auxiliary agents and that LNTs made from NaOH-lignin penetrate the cell nucleus. We further show that DNA can adsorb to LNTs. Consequently, exposure of HeLa cells to LNTs coated with DNA encoding the green fluorescent protein (GFP) leads to transfection and expression of GFP. The highest transfection efficiency was obtained with LNTs made from NaOH-lignin due to a combination of high DNA binding capacity and DNA delivery directly into the nucleus. These combined features of LNTs make LNTs attractive as smart delivery vehicles of DNA without the cytotoxicity associated with CNTs or the immunogenicity of viral vectors.

The hindgut lumen prokaryotic microbiota of the termite Reticulitermes flavipes and its responses to dietary lignocellulose composition.

Reticulitermes flavipes (Isoptera: Rhinotermitidae) is a highly eusocial insect that thrives on recalcitrant lignocellulosic diets through nutritional symbioses with gut-dwelling prokaryotes and eukaryotes. In the R. flavipes hindgut, there are up to 12 eukaryotic protozoan symbionts; the number of prokaryotic symbionts has been estimated in the hundreds. Despite its biological relevance, this diverse community, to date, has been investigated only by culture- and cloning-dependent methods. Moreover, it is unclear how termite gut microbiomes respond to diet changes and what roles they play in lignocellulose digestion. This study utilized high-throughput 454 pyrosequencing of 16S V5-V6 amplicons to sample the hindgut lumen prokaryotic microbiota of R. flavipes and to examine compositional changes in response to lignin-rich and lignin-poor cellulose diets after a 7-day feeding period. Of the ~475,000 high-quality reads that were obtained, 99.9% were annotated as bacteria and 0.11% as archaea. Major bacterial phyla included Spirochaetes (24.9%), Elusimicrobia (19.8%), Firmicutes (17.8%), Bacteroidetes (14.1%), Proteobacteria (11.4%), Fibrobacteres (5.8%), Verrucomicrobia (2.0%), Actinobacteria (1.4%) and Tenericutes (1.3%). The R. flavipes hindgut lumen prokaryotic microbiota was found to contain over 4761 species-level phylotypes. However, diet-dependent shifts were not statistically significant or uniform across colonies, suggesting significant environmental and/or host genetic impacts on colony-level microbiome composition. These results provide insights into termite gut microbiome diversity and suggest that (i) the prokaryotic gut microbiota is much more complex than previously estimated, and (ii) environment, founding reproductive pair effects and/or host genetics influence microbiome composition.

Influence of different cultivation methods on carbohydrate and lipid compositions and digestibility of energy of fruits and vegetables.

BACKGROUND: Environmental as well as cultivation factors may greatly influence the chemical composition of plants. The main factors affecting the chemical composition of foodstuff are level and type of fertilizer (conventional and organic cultivation systems), location or soil type, and year of harvest. Organic foods are defined as products that are produced under controlled cultivation conditions characterized by the absence of synthetic fertilisers and very restricted use of pesticides. Very limited information is available regarding the impact of organic cultivation systems on the composition of carbohydrates and fatty acids of fruits and vegetables. The objective was to investigate the influence of organic and conventional cultivation systems on the carbohydrate and fatty acid composition and digestibility of the energy of apple, carrot, kale, pea, potato, and rape seed oil. RESULTS: Carbohydrate and lignin values ranged from 584 g kg⁻¹ dry matter in kale to 910 g kg⁻¹ DM in potato, but with significant differences in the proportion of sugars, starch, non-starch polysaccharides, and lignin between the foodstuffs. Triacylglycerol was the major lipid class in pea, with 82% of total fatty acids, as opposed to apple, with only 35% of fatty acids of the ether extract. The most important factor influencing the digestibility of energy, and consequently faecal bulking, was the content of dietary fibre. CONCLUSION: The cultivation system had minor impact on the carbohydrate and lipid composition in the investigated foodstuffs or on the digestibility of energy when assessed in the rat model. Faecal bulking was related to dietary fibre in a linear fashion.

Determination of 67Zn distribution in navy bean (Phaseolus vulgaris L.) after foliar application of 67Zn-lignosulfonates using isotope pattern deconvolution.

The improvement of Zn fertilizers requires new techniques to evaluate their efficacy. In this paper, the (67)Zn stable isotope was used as tracer of several Zn-lignosulfonate complexes to study the foliar-applied Zn uptake and distribution behavior in the plant, compared with ZnEDTA. Navy bean plants ( Phaseolus vulgaris L.) were grown hydroponically in a Zn-free nutrient solution, and six modified lignosulfonates and EDTA complexed with (67)Zn were used in foliar application in the young leaves as Zn sources. Zinc isotopes in roots, stems, and sprayed and unsprayed leaves were determined by ICP-MS, and signal interferences caused by the compounds of the digested vegetal samples were corrected. The mathematical procedure of isotope pattern deconvolution allowed the minimization of the uncertainty in the measured molar fractions of Zn from fertilizer or from natural sources. Significant differences in Zn use and distribution were observed among the fertilizers when the calculated concentrations of Zn from the fertilizer were compared, whereas they were unnoticeable attending to the total Zn in plant tissues, usually determined at the conventional studies. By foliar spray, higher Zn uptake and mobilization to leaves and stems were achieved with (67)ZnEDTA than with (67)Zn-LS complexes. The ultrafiltered LS and phenolated LS showed slightly better ability to provide Zn to the bean plants than the other LS. The foliar-applied Zn use and distribution in the plant were related with the stability of the Zn-lignosulfonates complexes. Those presenting the lower stability versus pH, but the highest complexing capacity, were slightly more suitable to supply foliar-applied Zn to navy beans.