Influence of raw pea (Pisum sativum) or blue lupin seeds (Lupinus angustifolius) on the level of selected bioactive substances in pork meat

The study objective was to evaluate the impact of different contributions of pea (Pisum sativum) cultivar Hubal and blue lupin (Lupinus angustifolius) cultivar Regent on the level of selected bioactive substances in pork meat. 100 individuals three-breed cross piglets: ♀ (landrace × yorkshire) × ♂ duroc were used. Two experiments were performed, in which pea seeds (experiment I: E1 – 5.0% pea seeds; E2 – 10.0% pea seeds; E3 – 15.0% pea seeds; E4 -17.5% pea seeds) and blue lupin seeds (experiment II; D1 – 5.0% blue lupin seeds; D2 – 10.0% blue lupin seeds; D3 – 15.0% blue lupin seeds) were used instead of SBM-GM. In each of the experiments 50 animals were divided into 5 groups (control - C, and four experimental), placed in group pens, each for 10 individuals (sex ratio hogs : sows - 1:1). The animals were weighed and tagged before the experiments. The mean body weight of the pigs at experiment I and II commencement was: 26.7 and 33.5 kg, and at the end of the experiments: 122.0 and 124.0, respectively. In the first experiment (progressive pea contribution) the concentration of Carnosine was shown to be higher in E4 than E3 and C by 47.3% and 94.2%, respectively. In comparison with group C, the Q10 coenzyme content in groups E1, E2, E3 and E4 was lower by 40.9%; 56.8%; 40.9% and 65.9% respectively. In the second experiment (progressive lupin contribution) increased content of all of the investigated bioactive substances was recorded in groups D1-D3 vs C. Significant differences between groups C, D2, D4 for taurine (P≤0.05; P≤0.01) and creatine (P≤0.05) have been recorded. The content of bioactive substances in the longissimus lumborum muscle was significantly influenced by legumes, which increased the level of bioactive components of protein fraction. Therefore, it can be concluded, that pea (Pisum sativum) cultivar Hubal and blue lupine (Lupinus angustifolius) cultivar Regent are an alternative to SBM-GM, increasing the nutritionally valuable of pork meat.

A qPCR Assay for the Fast Detection and Quantification of Colletotrichum lupini

White lupin (Lupinus albus) represents an important legume crop in Europe and other parts of the world due to its high protein content and potential for low-input agriculture. However, most cultivars are susceptible to anthracnose caused by Colletotrichum lupini, a seed- and air-borne fungal pathogen that causes severe yield losses. The aim of this work was to develop a C. lupini-specific quantitative real-time TaqMan PCR assay that allows for quick and reliable detection and quantification of the pathogen in infected seed and plant material. Quantification of C. lupini DNA in dry seeds allowed us to distinguish infected and certified (non-infected) seed batches with DNA loads corresponding to the disease score index and yield of the mother plants. Additionally, C. lupini DNA could be detected in infected lupin shoots and close to the infection site, thereby allowing us to study the disease cycle of this hemibiotrophic pathogen. This qPCR assay provides a useful diagnostic tool to determine anthracnose infection levels of white lupin seeds and will facilitate the use of seed health assessments as a strategy to reduce the primary infection source and spread of this disease.

An Up-Scalable and Cost-Effective Methodology for Isolating a Polypeptide Matrix Metalloproteinase-9 Inhibitor from Lupinus albus Seeds

One of the most challenging problems with food-borne bioactive compounds is that there are commonly no cost-effective, generally recognized as safe (GRAS) methods for obtaining gram quantities of their purified forms. Here we aimed at developing a method to isolate deflamin, an oligomeric protein from lupin seeds with anti-inflammatory and anticancer activity through matrix metalloprotease (MMP)-9 inhibition. Our goal was to develop a GRAS method that could be easily up-scalable whilst maintaining deflamin’s activity. A sequential precipitation methodology was developed, using an aqueous extraction, followed by heat denaturation, acid precipitation and solubilization in ethanol. A final precipitation with 90% ethanol yielded a purified protein which was sequenced through mass spectrometry and tested for its MMP inhibitory activity using the Dye-quenched (DQ) gelatin assay and the standard wound healing assay in HT29 cells. The developed method yielded a purified oligomer, which represented 0.1% (w/w) of total dry seed weight and was positively confirmed to be deflamin. It further showed to effectively reduce MMP-9 gelatinolytic activity as well as colon cancer cell migration, hence corroborating the effectiveness of our method. Overall, this is the first reported method for isolating an MMP-9 inhibitor from legume seeds, which is up-scalable to an industrial level, in a cost-effective manner.

Untargeted Phytochemical Profile, Antioxidant Capacity and Enzyme Inhibitory Activity of Cultivated and Wild Lupin Seeds from Tunisia

Lupin seeds can represent a valuable source of phenolics and other antioxidant compounds. In this work, a comprehensive analysis of the phytochemical profile was performed on seeds from three Lupinus species, including one cultivar (Lupinus albus) and two wild accessions (Lupinus cossentinii and Lupinus luteus), collected from the northern region of Tunisia. Untargeted metabolomic profiling allowed to identify 249 compounds, with a great abundance of phenolics and alkaloids. In this regard, the species L. cossentinii showed the highest phenolic content, being 6.54 mg/g DW, followed by L. luteus (1.60 mg/g DW) and L. albus (1.14 mg/g DW). The in vitro antioxidant capacity measured by the ABTS assay on seed extracts ranged from 4.67 to 17.58 mg trolox equivalents (TE)/g, recording the highest values for L. albus and the lowest for L. luteus. The DPPH radical scavenging activity ranged from 0.39 to 3.50 mg TE/g. FRAP values varied between 4.11 and 5.75 mg TE/g. CUPRAC values for lupin seeds ranged from 7.20 to 8.95 mg TE/g, recording the highest for L. cossentinii. The results of phosphomolybdenum assay and metal chelation showed similarity between the three species of Lupinus. The acetylcholinesterase (AChE) inhibition activity was detected in each methanolic extract analyzed with similar results. Regarding the butyrylcholinesterase (BChE) enzyme, it was weakly inhibited by the Lupinus extracts; in particular, the highest activity values were recorded for L. albus (1.74 mg GALAE/g). Overall, our results showed that L. cossentinii was the most abundant source of polyphenols, consisting mainly in tyrosol equivalents (5.82 mg/g DW). Finally, significant correlations were outlined between the phenolic compounds and the in vitro biological activity measured, particularly when considering flavones, phenolic acids and lower-molecular-weight phenolics.

Quantification of Biologically Fixed Nitrogen by White Lupin (Lupins albus L.) and Its Subsequent Uptake by Winter Wheat Using the 15N Isotope Dilution Method

A field experiment was carried out in 2016–2018 in a white lupin (Lupinus albus L.)-winter wheat (Triticum aestivum cv. ‘Bogatka’) crop rotation. The aim of this study was to determine the amount of nitrogen (N) that was biologically fixed by the white lupin crop in the first year of the rotation and to estimate how much of this N was then taken up from the lupin residues by winter wheat in the second and third years of the rotation. Biologically fixed N was determined by the isotope-dilution method (ID15N) by applying 30 kg N ha−1 of 15N-labeled fertilizer (15NH4)2SO4 (containing 20.1 at.% 15N) to the white lupin and the reference plant spring wheat. The yields of white lupin seeds and crop residues were 3.92 t ha−1 and 4.30 t ha−1, respectively. The total amount of N in the white lupin biomass was 243.2 kg ha−1, which included 209.3 kg ha−1 in the seeds and 33.9 kg ha−1 in the residues. The 15N-labeled residue of white lupin was cut and ploughed into soil. Our results indicate that 111.2 kg N ha−1 was fixed from the atmosphere by the lupin plants, with 93.7 kg ha−1 found in the seeds and 17.5 kg ha−1 in the residues. In the second and third years of the rotation when winter wheat was cultivated, the plots were divided into two groups of subplots (1) without N-fertilization (control) and (2) with an application of 100 kg N ha−1. In the first year of winter wheat cultivation, 20.0% and 21.0% of N from the crop residues was taken up by the control and N-fertilization plots, respectively, while in the second year, uptake was lower at 7.12% and 9.27% in the control and N-fertilized plots, respectively.