Higher Protein Yield Research Articles

Trait Association Studies in Early Segregating Generations of Cowpea [Vigna unguiculata (L.) Walp

Background: Cowpea is a major pulse crop in India. Developing cowpea cultivars with high yield and sustainable protein content contributes to food and nutritional security. Hybridization followed by careful selection of genotypes from the early segregating populations may produce a final cultivar with high yield coupled with high protein content. Methods: The genetic material used in the present investigation were derived from crosses of three cultivars i.e., H10 (Anaswara x PKB 3) and H11 (Anaswara x PKB 4). The crosses were evaluated in the Department of Plant Breeding and Genetics, Kerala Agricultural University, Thrissur during cowpea season 2015-2017. Trait association based on correlation and path analysis was carried out with phenotypic data from F2 and F3 generations of the crosses. Result: In F2 generation of cross H10, number of pods per plant and number of seeds per pod displayed significant positive correlation with seed yield per plant. In F3 generation of cross H10, number of pods per plant and 100-seed weight per plant exhibited significant positive correlation with seed yield per plant. In cross H11, number of pods, pod length, single pod weight, number of seeds per pod and 100-seed weight revealed significant1 positive correlation with yield in segregating generations. On path analysis, various quantitative characters showed positive direct effects on seed yield. Selection of segregants based on the yield contributing characters is likely to be desirable for yield improvement in cowpea.

Physicochemical and functional properties of protein isolated from herring co-products; effects of catching season, pre-sorting, and co-product combination

Effect of catching season (spring vs fall), pre-sorting and selective recombination of different herring filleting co-products on protein yield during valorisation using the alkaline pH-shift technology was studied. Impacts of the pre-processing conditions on lipid oxidation, rheological, structural and functional properties of the proteins were also investigated. The sorted frame fraction resulted in the highest protein yield, myosin content, gel-forming capacity and gel whiteness. pH-shift processing triggered severe proteolytic degradation and lipid oxidation in the head fraction imposing a low-quality protein isolate. The unsorted co-products and the combinations head + frame and head + frame + tail gave protein isolates with gelation and oxidative quality being better than head but below the isolate from frame alone. The spring co-products produced protein isolates with better overall quality than the fall co-products. Altogether, the results revealed the advantage of sorting herring co-products, and the influence of season on protein extraction from herring co-products.

Coordinate Inheritance of Seed Isoflavone and Protein in Soybean

Soybean (Glycine max (L.) Merr.) is an important nutritional crop, not only as a rich source of protein and oil, but also for the production of isoflavones. There is a demand to breed soybean cultivars bearing consistently high protein, oil and isoflavone yields, yet this requires a clear heritable genetic relationship among isoflavone, protein and oil production. Here, two soybean genotypes contrasting in terms of protein, oil and isoflavone contents and their 185 F8:10 recombinant inbred lines (RILs) were employed to characterize seed protein, oil and isoflavone contents over two years of field trials. In this population, protein, but not oil, was significantly correlated with isoflavone content. A high-density genetic linkage map containing 3943 SNP markers identified through genotyping-by-sequencing (GBS) technology was constructed for further genetic analysis, whereby a total of 25 integrated isoflavone loci were identified, including qISO1, qISO 6.1, qISO 6.3 and qISO 6.4, which are newly identified QTLs. Two major QTLs identified in this study, qISO5 and qISO6.2, were fine-mapped for production of daidzein and genistein derivatives, as well as for glycitein derivatives, in the sequences between nucleotide positions 41042159 and 42098680 on chromosome 5 and between 18449510 and 19395795 on chromosome 6, which, respectively, explain 9.3–20.4% and 7.8–24.8% of the phenotypic variation in these traits. Further combination of qISO5 and qISO6.2 resulted in additive impacts on isoflavone production. Among the 13 QTLs linked with seed protein content in this study, three also colocated with QTLs for isoflavone content, indicating that seed isoflavone and protein content may be coordinately inherited. These results contribute to understanding the relationships between isoflavone and protein or oil content in soybean seeds. This knowledge could be valuable for soybean breeding programs aiming to combine consistently high isoflavone production with high protein or oil content.

CDC Reign Canada Prairie Spring Red wheat

CDC Reign, a hard red spring wheat ( Triticum aestivum L.), is adapted to the wheat growing regions of Western Canada and is eligible for the Canada Prairie Spring Red (CPSR) market class. This conventional height wheat combines high grain yield and grain protein concentration with strong straw, intermediate resistance to Fusarium head blight, and excellent milling properties. CDC Reign is resistant to leaf rust and moderately resistant to stem rust and demonstrated end-use quality suitable for the CPSR market class.

Reducing water uptake into BY-2 cells by systematically optimizing the cultivation parameters increases product yields achieved by transient expression in plant cell packs.

Plant-based production systems are inexpensive and easy to handle, allowing them to complement existing platforms for the production of protein-based vaccines, therapeutics and diagnostic reagents. However, screening product candidates in whole plants requires a large facility footprint and is challenging due to natural variations in recombinant protein accumulation. In contrast, plant cell packs (PCPs) allow more than 1000 samples to be screened per day in microtiter plates. PCPs enable rapid development cycles based on transient expression in as little as 3 days, and yield milligram quantities of product for initial quality assessment and functional testing. However, this requires high-level expression in BY-2 cells and consistent cell quality across batches. We therefore used a statistical design of experiments (DoE) approach to systematically assess factors that contribute to consistent high yields of recombinant proteins in PCPs. Specifically, we tested the osmolality, pH, carbon source, light source, and additives during cell cultivation, as well as cell and PCP harvest times. The careful adjustment of these factors increased overall productivity by approximately fourfold. Remarkably all cultivation conditions leading to high productivities during transient expression in PCPs were associated with a reduced water uptake into the central vacuole. The universal presence of a vacuole in plant cells indicates that our results should be transferrable to other cells lines. Our findings therefore support the broad application of PCPs for screening and product analysis during the development of protein-based pharmaceuticals and reagents in plants.

Cellulase pretreated palm decanter cake for feeding of black soldier fly larvae in triggering bioaccumulation of protein and lipid into biodiesel productions

For every ton of palm oil produced, approximately 0.2 tons of palm decanter cake (PDC) waste is generated. Hence, there is a huge opportunity to valorize this organic waste, i.e., via the deployment of black soldier fly larvae (BSFL), which has been widely employed to convert various organic wastes into larval proteins and lipids. However, the PDC is mainly made up of lignocellulosic materials that are hard to digest by the BSFL. Therefore, this work attempted to grow BSFL in cellulase-pretreated PDC; thereby, providing an alternative solution to manage PDC waste. Results had shown that the cellulase pretreatment was effective in breaking down cellulose into glucose molecules, especially with higher cellulase dosage and longer treatment duration of up until 48 h. Subsequently, the maximum BSFL growth was found at 6.56 ± 2.69 mg/larva when being fed with PDC pre-treated by 1.0 wt% of cellulase for 72 h; which was about 4 mg higher than the controlled larva. Using a similar substrate, the highest protein yield and lipid yield from BSFL were attained at 1.63 ± 0.11 mg/larva (22.4 wt%) and 5.12 ± 1.01 mg/larva (69.9 wt%), respectively. In terms of biodiesel quality, a huge presence of saturated fatty acids had made the BSFL-based biodiesel oxidatively stable.

High-throughput strategy for identification of Mycobacterium tuberculosis membrane protein expression conditions using folding reporter GFP

Mycobacterium tuberculosis membrane protein biochemistry and structural biology studies are often hampered by challenges in protein expression and selection for well-expressing protein candidates, suitable for further investigation. Here we present a folding reporter GFP (frGFP) assay, adapted for M. tuberculosis membrane protein screening in Escherichia coli Rosetta 2 (DE3) and Mycobacterium smegmatis mc24517. This method allows protein expression condition screening for multiple protein targets simultaneously by monitoring frGFP fluorescence in growing cells. We discuss the impact of common protein expression conditions on 42 essential M. tuberculosis H37Rv helical transmembrane proteins and establish the grounds for their further analysis. We have found that the basal expression of the lac operon in the T7-promoter expression system generally leads to high recombinant protein yield in M. smegmatis, and we suggest that a screening condition without the inducer is included in routine protein expression tests. In addition to the general observations, we describe conditions allowing high-level expression of more than 25 essential M. tuberculosis membrane proteins, containing 2 to 13 transmembrane helices. We hope that these findings will stimulate M. tuberculosis membrane protein research and aid the efforts in drug development against tuberculosis.

Utilization of Recombinant Baculovirus Expression System to Produce the RBD Domain of SARS-CoV-2 Spike Protein.

Continuous outbreaks of viral diseases in humans facilitates a need for the rapid development of viral test kits and vaccines. These require expression systems to produce a pure and high yield of target viral proteins. We utilized a baculovirus–silkworm expression system to produce the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. First, we had to develop a strategy for constructing a recombinant baculovirus for RBD expression. For this, the coding region of the Bombyx mori cypovirus (BmCPV) polyhedron was assembled with the Bombyx mori nuclear polyhedrosis virus (BmNPV) promoter. We demonstrated that the recombinant baculovirus has the ability to form polyhedrons within host silkworm cells. In addition, the encapsulated BVs are able to infect silkworms by ingestion and induce foreign protein expression. In this way, we utilized this novel system to obtain a high yield of the target foreign protein, the RBD of the SARS-CoV-2 S protein. However, the viral infection rate of our recombinant BV needs to be improved. Our study shed light on developing a highly efficient expression system for the production of antigens and subsequent immunoassays and vaccines.

The Quality and Use Value of the False Flax (Camelina sativa [L.] Crantz)

Summary Alternative plant cultures with favorable agronomic characteristics are a great source of refined edible oil and renewable industrial oil products. False flax stands out as a species with modest growing requirements, short vegetation and diverse use. Accordingly, the breeding program of the Institute of Field and Vegetable Crops in Novi Sad created two cultivars of false flax, NS Zlatka and NS Slatka, which were the research material of this study. The use and production value of these false flax cultivars were determined by examining important seed quality characteristics: oil content, oil yield, protein content and protein yield. The highest oil content was obtained in NS Slatka (45.90%) in 2016, while the highest protein content was 30.30%. The highest oil yield (801 kg/ha in 2017) and the highest protein yield (424 kg/ha in 2017) were detected in NS Zlatka. The data obtained from the field and laboratory were analyzed by analysis of variance, Duncan test and Spearman's correlation coefficient. Statistically significant differences were identified between the cultivars, and strong environmental influence was affirmed. By examining the correlations of the analyzed features, statistically significant correlations were found. The most common fatty acids included linolenic acid (33.80%), linoleic acid (18.81%), oleic acid (15.99%), eicosenoic acid (14.22%) and erucic acid (2.90%). The results obtained are applicable in further breeding program of false flax. They are of special importance in increasing the use value of false flax owing to the oil and meal high quality and potential for even more diverse use.

Effect of Crop Establishment Methods and Microbial Inoculations on Augmenting the Energy Efficiency and Nutritional Status of Rice and Wheat in Cropping System Mode

A field experiment was conducted for two consecutive years with the aim to quantify the role of different nutrient management variables such as microbial inoculation, zinc (Zn) fertilization and optimal and sub-optimal fertilization of nitrogen and phosphorus on the energetic and nutritional status of the rice–wheat cropping system (RWCS). The said nutrient management variables were applied over six different crop establishment methods (CEMs) in RWCS viz. puddled transplanted rice (PTR), system of rice intensification (SRI) and aerobic rice system (ARS) in rice and conventional drill-sown wheat (CDW), system of wheat intensification (SWI) and zero-tillage wheat (ZTW) in wheat. Two microbial consortia viz. Anabaena sp. (CR1) + Providencia sp. (PR3) consortia (MC1) and Anabaena-Pseudomonas biofilmed formulations (MC2) were used in this study, while recommended dose of nitrogen (N) and phosphorus (P) (RDN) (120 kg N ha−1 and 25.8 kg P ha−1), 75% RDN and Zn fertilization (soil applied 5 kg Zn ha−1 through zinc sulphate heptahydrate) were the other variables. The contribution of microbial consortia, Zn fertilization and RDN (over 75% RDN) to net energy production of RWCS was 12.9–16.1 × 103 MJ ha−1, 10.1–11.0 × 103 MJ ha−1 and 11.7–15.3 × 103 MJ ha−1. Among the CEMs, the highest gross and net energy production was recorded in ARS–ZTW with lowest energy required for production of one tonne of system yield (2366–2523 MJ). The system protein yield varies from 494.1 to 957.7 kg ha−1 with highest protein yield in 75% RDN + MC2 + Zn applied ARS–ZTW. Among micronutrients, the uptake of Zn and iron (Fe) is sensitive to all studied variables, while manganese (Mn) and cupper (Cu) uptake was found significantly affected by CEMs alone. The combination of 75% RDN + MC2 + Zn in ARS–ZTW was found superior in all respects with 288.3 and 286.9 MJ ha−1 net energy production and 2320 and 2473 MJ energy required for production of one tonne system yield in the first and second year of study, respectively.

High-Yield Production of Catalytically Active Regulatory [NiFe]-Hydrogenase From Cupriavidus necator in Escherichia coli.

Hydrogenases are biotechnologically relevant metalloenzymes that catalyze the reversible conversion of molecular hydrogen into protons and electrons. The O2-tolerant [NiFe]-hydrogenases from Cupriavidus necator (formerly Ralstonia eutropha) are of particular interest as they maintain catalysis even in the presence of molecular oxygen. However, to meet the demands of biotechnological applications and scientific research, a heterologous production strategy is required to overcome the low production yields in their native host. We have previously used the regulatory hydrogenase (RH) from C. necator as a model for the development of such a heterologous hydrogenase production process in E. coli. Although high protein yields were obtained, the purified enzyme was inactive due to the lack of the catalytic center, which contains an inorganic nickel-iron cofactor. In the present study, we significantly improved the production process to obtain catalytically active RH. We optimized important factors such as O2 content, metal availability, production temperature and time as well as the co-expression of RH-specific maturase genes. The RH was successfully matured during aerobic cultivation of E. coli by co-production of seven hydrogenase-specific maturases and a nickel permease, which was confirmed by activity measurements and spectroscopic investigations of the purified enzyme. The improved production conditions resulted in a high yield of about 80 mg L–1 of catalytically active RH and an up to 160-fold space-time yield in E. coli compared to that in the native host C. necator [<0.1 U (L d) –1]. Our strategy has important implications for the use of E. coli K-12 and B strains in the recombinant production of complex metalloenzymes, and provides a blueprint for the production of catalytically active [NiFe]-hydrogenases in biotechnologically relevant quantities.

A CHO-Based Cell-Free Dual Fluorescence Reporter System for the Straightforward Assessment of Amber Suppression and scFv Functionality.

Incorporation of noncanonical amino acids (ncAAs) with bioorthogonal reactive groups by amber suppression allows the generation of synthetic proteins with desired novel properties. Such modified molecules are in high demand for basic research and therapeutic applications such as cancer treatment and in vivo imaging. The positioning of the ncAA-responsive codon within the protein’s coding sequence is critical in order to maintain protein function, achieve high yields of ncAA-containing protein, and allow effective conjugation. Cell-free ncAA incorporation is of particular interest due to the open nature of cell-free systems and their concurrent ease of manipulation. In this study, we report a straightforward workflow to inquire ncAA positions in regard to incorporation efficiency and protein functionality in a Chinese hamster ovary (CHO) cell-free system. As a model, the well-established orthogonal translation components Escherichia coli tyrosyl-tRNA synthetase (TyrRS) and tRNATyrCUA were used to site-specifically incorporate the ncAA p-azido-l-phenylalanine (AzF) in response to UAG codons. A total of seven ncAA sites within an anti-epidermal growth factor receptor (EGFR) single-chain variable fragment (scFv) N-terminally fused to the red fluorescent protein mRFP1 and C-terminally fused to the green fluorescent protein sfGFP were investigated for ncAA incorporation efficiency and impact on antigen binding. The characterized cell-free dual fluorescence reporter system allows screening for ncAA incorporation sites with high incorporation efficiency that maintain protein activity. It is parallelizable, scalable, and easy to operate. We propose that the established CHO-based cell-free dual fluorescence reporter system can be of particular interest for the development of antibody-drug conjugates (ADCs).

Physical Properties of Extrudates with Fibrous Structures Made of Faba Bean Protein Ingredients Using High Moisture Extrusion.

Faba bean is a potential ingredient due to its high protein yield and its possible cultivation in colder climate regions. In this study, meat analogues made from faba bean protein isolate (FPI) and concentrate (FPC) blends were produced using high moisture extrusion. The aim of this study was to investigate the effect of the FPI content (FPIc), feed water content (FWC), and temperature of the long cooling die (LT) during extrusion on the mechanical and physicochemical properties as well as on the structure of the meat analogues. Increased FPIc resulted in higher values in hardness, gumminess, chewiness, and cutting strengths as well as in darker colour and decreased water absorption capacity. The effect of increased FWC on these properties was weaker and the opposite. Images from microtomography revealed that higher FPIc led to a less organised fibrous structure. In conclusion, fibrous structures can be achieved by utilising a mixture of faba bean protein ingredients, and a higher FPC content seemed to promote fibre formation in the meat analogue.

Characterizing Extracellular Vesicles and Particles Derived from Skeletal Muscle Myoblasts and Myotubes and the Effect of Acute Contractile Activity.

Extracellular vesicles (EVs), released from all cells, are essential to cellular communication and contain biomolecular cargo that can affect recipient cell function. Studies on the effects of contractile activity (exercise) on EVs usually rely on plasma/serum-based assessments, which contain EVs from many different cells. To specifically characterize skeletal muscle–derived vesicles and the effect of acute contractile activity, we used an in vitro model where C2C12 mouse myoblasts were differentiated to form myotubes. EVs were isolated from conditioned media from muscle cells at pre-differentiation (myoblasts) and post-differentiation (myotubes) and also from acutely stimulated myotubes (1 h @ 14 V, C-Pace EM, IonOptix, Westwood, MA, USA) using total exosome isolation reagent (TEI, ThermoFisher (Waltham, MA, USA), referred to as extracellular particles [EPs]) and differential ultracentrifugation (dUC; EVs). Myotube-EPs (~98 nm) were 41% smaller than myoblast-EPs (~167 nm, p < 0.001, n = 8–10). Two-way ANOVA showed a significant main effect for the size distribution of myotube vs. myoblast-EPs (p < 0.01, n = 10–13). In comparison, myoblast-EPs displayed a bimodal size distribution profile with peaks at <200 nm and 400–600, whereas myotube-Eps were largely 50–300 nm in size. Total protein yield from myotube-EPs was nearly 15-fold higher than from the myoblast-EPs, (p < 0.001 n = 6–9). Similar biophysical characteristics were observed when EVs were isolated using dUC: myotube-EVs (~195 nm) remained 41% smaller in average size than myoblast-EVs (~330 nm, p = 0.07, n = 4–6) and had comparable size distribution profiles to EPs isolated via TEI. Myotube-EVs also had 4.7-fold higher protein yield vs. myoblast EVs (p < 0.05, n = 4–6). Myotube-EPs exhibited significantly decreased expression of exosomal marker proteins TSG101, CD63, ALIX and CD81 compared with myoblast-EPs (p < 0.05, n = 7–12). Conversely, microvesicle marker ARF6 and lipoprotein marker APO-A1 were only found in the myotube-EPs (p < 0.05, n = 4–12). There was no effect of acute stimulation on myotube-EP biophysical characteristics (n = 7) or on the expression of TSG101, ARF6 or CD81 (n = 5–6). Myoblasts treated with control or acute stimulation–derived EPs (13 µg/well) for 48 h and 72 h showed no changes in mitochondrial mass (MitoTracker Red, ThermoFisher, Waltham, MA, USA), cell viability or cell count (n = 3–4). Myoblasts treated with EP-depleted media (72 h) exhibited ~90% lower cell counts (p < 0.01, n = 3). Our data show that EVs differed in size, distribution, protein yield and expression of subtype markers pre vs. post skeletal muscle–differentiation into myotubes. There was no effect of acute stimulation on biophysical profile or protein markers in EPs. Acute stimulation–derived EPs did not alter mitochondrial mass or cell count/viability. Further investigation into the effects of chronic contractile activity on the biophysical characteristics and cargo of skeletal muscle–specific EVs are warranted.

Growth Characteristics of Chlorella sorokiniana in a Photobioreactor during the Utilization of Different Forms of Nitrogen at Various Temperatures.

The cultivation of microalgae requires the selection of optimal parameters. In this work, the effect of various forms of nitrogen on the growth and productivity of Chlorella sorokiniana AM-02 when cultivated at different temperatures was evaluated. Regardless of the temperature conditions, the highest specific growth rate of 1.26 day−1 was observed in modified Bold’s basal medium (BBM) with NH4+ as a nitrogen source, while the highest specific growth rate in BBM with NO3− as a nitrogen source achieved only 1.07 day−1. Moreover, C. sorokiniana grew well in medium based on anaerobic digester effluent (ADE; after anaerobic digestion of chicken/cow manure) with the highest growth rate being 0.92 day−1. The accumulation of proteins in algal cells was comparable in all experiments and reached a maximum of 42% of dry weight. The biomass productivity reached 0.41–0.50 g L−1 day−1 when cultivated in BBM, whereas biomass productivity of 0.32–0.35 g L−1 day−1 was obtained in ADE-based medium. The results, based on a bacterial 16S rRNA gene sequencing approach, revealed the growth of various bacterial species in ADE-based medium in the presence of algal cells (their abundance varied depending on the temperature regimen). The results indicate that biomass from C. sorokiniana AM-02 may be sustainable for animal feed production considering the high protein yields.

Processing Condition and Properties of Protein Isolates from Mung Bean (Vigna radiata) and Quinoa (Chenopodium quinoa Willd)

Mung bean and quinoa are good sources of protein from legumes and pseudo cereals respectively. Both sources have relatively high protein content and acceptable amino acids composition. The potential and application of mung bean and quinoa as protein isolates have been discovered in some studies. Several isolation techniques such as wet fractionation (acid-base extraction), micellization, dry fractionation, aqueous separation or combination of some techniques can be used to isolate protein. This review summarized different methods and processing conditions applied during production of mung bean and quinoa protein isolates. Extracting protein from different sources may require different conditions, which results in different compositions and functional properties. Acid-base extraction is the most common method applied in mung bean and quinoa and results in high protein purity and yields. Micellization is an alternative method used in mung bean to produce higher protein content. Dry fractionation is a sustainable option used in quinoa to concentrate protein fractions. Purification methods such as ultrafiltration and aqueous phase separation can be used. Different methods and processing conditions affect functional properties, including solubility, water and oil absorption, emulsification, foaming, thermal properties of isolates, which also affect the suitable application for isolates.&#x0D; Keywords: Protein isolates; Quinoa; Mung bean; Wet fractionation; Dry fractionation

Wheat–Lentil Mixed Cropping System Productivity Under Varied Irrigation Levels

The study on mixed cropping of lentil and wheat was conducted under three levels of irrigation. Wheat at 10, 15, 20, 25, 30, 35, 40, 45 and 50% of the normal seed rates were mixed with full seed rate of lentil to find out a suitable mixing rate of wheat for maximum total yield from unit land. The study was conducted in a split plot design with three replications. Lentil yields obtained with single and two irrigations were identical and higher than with no irrigation. The highest wheat yield was obtained with two irrigations. Total yield was the highest with application of two irrigations. Mixing different proportions of wheat with lentil produced 95-171% additional yield over sole cropping of lentil. Mixing of wheat at 25-35% with lentil was found to be judicious for obtaining maximum total yields. The highest protein yield was obtained with 30-35% and 35% mixing rates of wheat at zero and single irrigation levels. Growing wheat with lentil resulted in increased protein yield in wheat than sole cropping. The highest land equivalent ratio (LER) values were obtained when 30-35% of wheat seed rate were mixed with lentil. Application of two irrigations resulted in high LER values. It is concluded that use of 35% of wheat seed rate along with full seed rate of lentil under two levels of irrigations would be the best mixing ratio for obtaining the highest wheat and total grain yield. Ann. Bangladesh Agric. (2021) 25 (1) : 1-10

Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels.

Epizootic haemorrhagic disease virus (EHDV) is a member of the Orbivirus genus in the Reoviridae family, and it is the etiological agent of an arthropod-transmitted disease that affects domestic and wild ruminants. Due to its significant economic impact, many attempts have been done in order to develop diagnostic immunoassays mainly based on the use of the viral protein 7 (VP7), that is, the immunodominant serogroup-specific antigen. In this work, a recombinant VP7 (recVP7) of EHDV serotype 2 was produced in a baculovirus system, and after purification using ion metal affinity chromatography, we obtained a high yield of recombinant protein characterized by a high degree of purity. We used the purified recVP7 as reagent to develop a competitive enzyme-linked immunoassay (c-ELISA), and we tested the presence of EHDV antibodies in 185 dromedary camel serum samples. The c-ELISA showed good performance parameters in recognising positive sera of naturally EHDV-infected dromedary camels; in particular, our developed test reached 85.7% of sensitivity, 98.1% of specificity, 93% of accuracy, and a high agreement value with results obtained by the commercial ELISA kit (Cohen's kappa value of 0.85) that we adopted as the reference method. This c-ELISA could be a useful screening test to monitor the virus spread in camels that are sentinel animals for endemic areas of disease.

Novel application of ultrasound and microwave-assisted methods for aqueous extraction of coconut oil and proteins.

Alternative methods for wet extraction of coconut oil and protein assisted by ultrasound or microwave were developed and compared. Coconut milk was prepared by milling the pulp (5:1 water to coconut pulp ratio), further destabilised at pH 4 and centrifuged to obtain the cream and cream protein fractions (control process). Microwave-assisted treatment applied in milk (1min, 3 pulses of 20s; 2.5GHz; 4.31kW/kg by pulse) generated a significant increase in cream obtained, and in the coconut oil extraction yield (~ 20%) compared to its control. The ultrasound-assisted treatment (2.5min; 24kHz; 0.573kW/kg, 6.85W/cm2) also improved oil extraction (10-16%). Moreover, a higher protein yield was achieved in ultrasound treated samples when compared to their control (49.6-86.1%). Large particles of 11m , probably aggregates of particles, and smaller particles of 3.6m , were detected in coconut milk, which were reduced by ultrasound effect. Alternative treatments caused a greater liberation of total phenols in coconut cream. Coconut proteins in water (0.1%) showed high negative electrokinetic potential. The surface pressure of coconut proteins at the air/water interface was not modified by assisted treatments.

Historical grain yield genetic gains in Norwegian spring wheat under contrasting fertilization regimes

AbstractSpring wheat is currently dominating wheat production in Norway. The introduction of combine harvesting in the 1950s spurred breeding efforts to improve lodging and preharvest sprouting resistance, and to integrate good breadmaking quality in the locally adapted germplasm. Release of landmark cultivars Runar and Reno in the 1970s revitalized the country's production and marked the onset of modern wheat cultivation in Norway. Since that time, new cultivars have been developed but little is known about the genetic basis of the achieved yield gains. We collected 21 representative cultivars released since 1972 in Norway and tested them in a multiyear field trial including two fertilization rates: 75 and 150 kg ha–1 N. We assessed grain yield, plant height, heading, maturity, length of grain filling period, grain protein content, protein yield, aboveground biomass, harvest index, grain weight, test weight, grains per spike, grains per square meter, and spikes per square meter and their response to fertilization. We document an annual increase in grain yield of 17.8 kg ha–1 (0.34%), at both rates of N fertilization. None of the traits exhibited significant genotype × management interaction. Wheat breeding has led to the development of higher‐yielding cultivars with higher protein yield that mature later, have a prolonged grain‐filling period, and produce more grains per spike and grains per unit area.