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The increased fish consumption by the growing human population in the world translates into an increase in fish waste. The reintroduction of these fish by-products into food and feed chains presents economic benefits and contributes to counteracting their negative environmental impact. Under this context, the present study aimed to evaluate the effects of the dietary inclusion of fish hydrolysate and oil obtained from fish waste (experimental diet) in substitution of shrimp hydrolysate and salmon oil (control diet) mainly imported from third countries on palatability, apparent total tract digestibility, fecal characteristics and metabolites, blood fatty acid profile, flatulence, and coat quality of adult dogs. A two-bowl test was performed to evaluate palatability by the pairwise comparison between the two diets. A feeding trial was conducted according to a crossover design with two diets (control and experimental diets), six adult Beagle dogs per diet, and two periods of 6 weeks each. The replacement of shrimp hydrolysate and salmon oil with fish hydrolysate and oil did not affect the first diet approach and taste, as well as the intake ratio. Generally, the digestibility of dry matter, nutrients, and energy was not affected by diet, but the intake of digestible crude protein (CP) and ether extract was higher, respectively, with the control and the experimental diet. The higher intake of eicosapentaenoic acid and docosahexaenoic acid with the experimental diet was reflected in a higher content of these long-chain polyunsaturated fatty acids and the omega-3 index of red blood cells, but it did not affect coat quality. The significantly higher intake of digestible CP with the control diet might have contributed to the higher fecal ammonia-N and valerate concentrations. Daily fecal output and characteristics were similar between diets. Overall, results suggest that fish hydrolysate and oil from the agrifood industry might constitute sustainable functional ingredients for dog feeding while adding value for wild fisheries, aquaculture, and fish farming under a circular economy approach and reducing dependence on imports from third countries with a high carbon footprint. [ABSTRACT FROM AUTHOR]

The in vitro rumen batch technique is widely used for screening novel feed sources; however, it remains unclear to what extent the in vitro fermentability of non-conventional feed sources is affected by non-adapted ruminal inocula. Thus, in this study, we evaluated the effects of distinct ruminal inocula on the in vitro fermentation parameters of a sustainable non-conventional feed, a commercially available algal blend composed of microalgae (Chlorella vulgaris and Nannochloropsis oceanica) and seaweeds (Ulva sp. and Gracilaria gracilis). First, four late-lactation Holstein cows were fed four forage-based diets varying only in the proportions of basal forage (100% corn silage, 70% corn silage and 30% haylage, 30% corn silage and 70% haylage, and 100% haylage) in a 4 x 4 Latin square design with the last square omitted. After 3 weeks of adaptation, haylage-based diets resulted in ruminal fermentation parameters distinct from those promoted by corn silage-based diets, as reflected in increased pH, ammonia-N contents, and acetate proportions. Individual ruminal fluids derived from each of the four diets were further used as inocula in in vitro incubations. Here, a 1:1 mixture of corn silage and haylage was supplemented with 0, 5, 10, or 15% algal blend and incubated with each inoculum for 24 h in a 4 x 4 factorial design. Total gas and methane production decreased with inocula from cows fed haylage-based diets and with increasing algal blend supplementation levels. The fermentation pH increased and the ammonia-N contents decreased with inocula from cows fed haylage-based diets; however, these parameters were not affected by algal blend inclusion levels. The interaction between the ruminal inoculum source and the algal blend supplementation level affected the total volatile fatty acids (VFA) and the proportions of most individual VFA. Total VFA production decreased with increasing algal supplementation levels, particularly with inocula from cows fed 30% corn silage and 70% haylage; the acetate, propionate, and valerate proportions were only affected by algal blend levels under incubation with 100% corn silage inocula. Overall, our findings highlight the importance of the ruminal inoculum source when assessing the fermentability of non-conventional feed as well as the potential of the algal blend as a natural modulator of ruminal fermentation. [ABSTRACT FROM AUTHOR]

Background. The awareness of environmental and socio-economic impacts caused by greenhouse gas emissions from the livestock sector leverages the adoption of strategies to counteract it. Feed supplements can play an important role in the reduction of the main greenhouse gas produced by ruminants methane (CH4). In this context, this study aims to assess the effect of two biochar sources and inclusion levels on rumen fermentation parameters in vitro. Methods. Two sources of biochar (agro-forestry residues, AFB, and potato peel, PPB) were added at two levels (5 and 10%, dry matter (DM) basis) to two basal substrates (haylage and corn silage) and incubated 24-h with rumen inocula to assess the effects on CH4 production and main rumen fermentation parameters in vitro. Results. AFB and PPB were obtained at different carbonization conditions resulting in different apparent surface areas, ash content, pH at the point of zero charge (pHpzc), and elemental analysis. Relative to control (0% biochar), biochar supplementation kept unaffected total gas production and yield (mL and mL/g DM, pD0:140 and pD0:240, respectively) and fermentation pH (p D 0:666), increased CH4 production and yield (mL and mL/g DM, respectively, pD0:001) and ammonia-N (NH3-N, pD0:040), and decreased total volatile fatty acids (VFA) production (p < 0:001) and H2 generated and consumed (p 0:001). Biochar sources and inclusion levels had no negative effect on most of the fermentation parameters and efficiency. Acetic:propionic acid ratio (pD0:048) and H2 consumed (pD0:019) were lower with AFB inclusion when compared to PPB. Biochar inclusion at 10% reduced H2 consumed (p < 0:001) and tended to reduce total gas production (pD0:055). Total VFA production (pD0:019), acetic acid proportion (pD0:011) and H2 generated (pD0:048) were the lowest with AFB supplemented at 10%, no differences being observed among the other treatments. The basal substrate affected most fermentation parameters independently of biochar source and level used. Discussion. Biochar supplementation increasedNH3-N content, iso-butyric, iso-valeric and valeric acid proportions, and decreased VFA production suggesting a reduced energy supply for microbial growth, higher proteolysis and deamination of substrate N, and a decrease of NH3-N incorporation into microbial protein. No interaction was found between substrate and biochar source or level on any of the parameters measured. Although AFB and PPB had different textural and compositional characteristics, their effects on the rumen fermentation parameters were similar, the only observed effects being due to AFB included at 10%. Biochar supplementation promoted CH4 production regardless of the source and inclusion level, suggesting that there may be other effects beyond biomass and temperature of production of biochar, highlighting the need to consider other characteristics to better identify the mechanism by which biochar may influence CH4 production. [ABSTRACT FROM AUTHOR]

Chemical analyses, enzymatic and Tilley & Terry digestibility were used to describe the feeding values of eight alfalfa samples. Six fresh alfalfa samples of different successive cuts and two alfalfa hays of two successive cuts of local varieties grown in Albania were used in this study. All samples were analysed for their chemical content of dry matter (DM), crude protein (CP), ash, crude fat (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF), acid detergent lignin (ADL), and digestibility of organic matter (OMD). Fresh alfalfa samples had significant variation on chemical content for main Weende parameters. Early and late cut alfalfas have lower values for NDF, ADF and ADL content compering to medium cut. The values for CP of fresh alfalfas ranged from (17.0-22.7%) and for alfalfa hays (12.2-19.7%), the content of CF ranged from (28.2-40.0%) for fresh alfalfas and (26.8-29.6%) for hays. Ash content varied from (8.60-10.8%) and (8.90-10.2%), respectively for fresh and hays alfalfas. The NDF, ADF and ADL content of alfalfas varied respectively (39.6-52.4%), (30.5- 43.1 %), (5.70-10.3%) for fresh alfalfas and (38.0-47.5%), (31.4-33.4%), (6.40-7.60%) for hays. The OMD digestibility values determined with Tilley and Terry method for all feeds included in the study resulted higher than OMD determined with enzymatic method respectively (66.9 vs 51.3%). The results of OMD determined by two "in vitro" methods were strongly correlated. According to R 2-value (0.89) the OMD determined by Tilley and Terry method could be predicted from enzymatic test as most convenient one since it does not need animals. [ABSTRACT FROM AUTHOR]

Omega-3 fatty acids (O3FA) are essential nutrients that play a crucial role in maintaining human and animal health. They are known for their numerous health claims, including cardiovascular benefits, contributing to both the prevention and treatment of immunological, neurological, reproductive, and cardiovascular complications, and supporting overall well-being. Fish, especially oily fish, comprise rich source of O3FA. In the fish industry, significant amounts of by-products and waste are generated during processing which are often discarded or used for lower-value applications. However, there is recognition of the potential value of extracting O3FA from these by-products. Various extraction techniques can be used, but the goal is to efficiently extract and concentrate the O3FA while minimizing the loss of nutritional value. To prevent oxidation and maintain the stability of O3FA, natural antioxidants can be added. Antioxidants like polyphenolic compounds and plant extracts help to protect the O3FA from degradation caused by exposure to oxygen, light, and heat. By stabilizing the O3FA, the shelf life and nutritional value of the extracted product can be extended. In summary, this work presents a forward-looking strategy for transforming fish by-products into high-quality oils, which hold great potential for application in food and feed. [ABSTRACT FROM AUTHOR]

Seasonal and daily variations of gaseous emissions from naturally ventilated dairy cattle barns are important figures for the establishment of effective and specific mitigation plans. The present study aimed to measure methane (CH 4 ) and ammonia (NH 3 ) emissions in three naturally ventilated dairy cattle barns covering the four seasons for two consecutive years. In each barn, air samples from five indoor locations were drawn by a multipoint sampler to a photoacoustic infrared multigas monitor, along with temperature and relative humidity. Milk production data were also recorded. Results showed seasonal differences for CH 4 and NH 3 emissions in the three barns with no clear trends within years. Globally, diel CH 4 emissions increased in the daytime with high intra-hour variability. The average hourly CH 4 emissions (g h -1 livestock unit -1 (LU)) varied from 8.1 to 11.2 and 6.2 to 20.3 in the dairy barn 1, from 10.1 to 31.4 and 10.9 to 22.8 in the dairy barn 2, and from 1.5 to 8.2 and 13.1 to 22.1 in the dairy barn 3, respectively, in years 1 and 2. Diel NH 3 emissions highly varied within hours and increased in the daytime. The average hourly NH 3 emissions (g h -1  LU -1 ) varied from 0.78 to 1.56 and 0.50 to 1.38 in the dairy barn 1, from 1.04 to 3.40 and 0.93 to 1.98 in the dairy barn 2, and from 0.66 to 1.32 and 1.67 to 1.73 in the dairy barn 3, respectively, in years 1 and 2. Moreover, the emission factors of CH 4 and NH 3 were 309.5 and 30.6 (g day -1  LU -1 ), respectively, for naturally ventilated dairy cattle barns. Overall, this study provided a detailed characterization of seasonal and daily gaseous emissions variations highlighting the need for future longitudinal emission studies and identifying an opportunity to better adequate the existing mitigation strategies according to season and daytime., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The identification of crops that simultaneously contribute to the global protein supply and mitigate the effects of climate change is an urgent matter. Lupins are well adapted to nutrient-poor or contaminated soils, tolerate various abiotic stresses, and present relevant traits for acting as ecosystem engineers. Lupins are best studied for their seeds, but their full foraging potential needs further evaluation. This study evaluated the effects of location and sowing date on forage production, proximate composition, and the detailed mineral and alkaloid profiles of three species of Lupinus ( L. albus cv. Estoril, L. angustifolius cv. Tango, and L. luteus cv. Cardiga). Sowing date and location and their interaction with the plant species significantly affected the vast majority of measured parameters, emphasizing the effects of climate and soil conditions on these crops. The relatively high crude protein and in vitro digestibility support the potential of the lupin species studied as sustainable forage protein sources in diets for ruminant animals. The content of individual essential macro and trace elements was below the maximum tolerable levels for cattle and sheep. Lupanine, smipine, and sparteine were the most abundant quinolizidine alkaloids in L. albus cv. Estoril, lupanine, and sparteine in L. angustifolius cv. Tango, and lupinine, gramine, ammodendrine, and sparteine in L. luteus cv. Cardiga. Based on the maximum tolerable levels of total quinolizidine alkaloid intake, the dietary inclusion of forages of L. albus cv. Estoril and L . angustifolius cv. Tango does not pose a risk to the animals, but the high alkaloid content of L. luteus cv. Cardiga may compromise its utilization at high levels in the diet. Overall, the results reveal a high potential for lupins as protein forage sources well adapted to temperate regions and soils with lower fertility, with a relevant impact on livestock sustainability in a climate change era., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The production of Lupinus seeds for food and feed is increasing worldwide, which results in large amounts of post-harvest biomass residues, considered of low value and left in the field to be burned or incorporated in the soil. To valorize these agricultural wastes, this work aimed to assess their potential as an alternative feed for ruminants. Thus, the production yield, nutritive value, and alkaloid content of straws and pod shells from three native European Lupinus species, L. albus 'Estoril' (white), L. angustifolius 'Tango' (narrow-leafed), and L. luteus 'Cardiga' (yellow), cultivated in two locations, were evaluated. The dry matter (DM) yield of straws and pod shells were the highest for L. albus 'Estoril' (4.10 t ha -1 ) and the lowest for L. angustifolius 'Tango' (1.78 t ha -1 ), suggesting a poor adaptation of narrow-leafed lupin to the particularly dry and warm agronomic year. Despite species-specific differences, lupin biomass residues presented higher crude protein (53.0-68.9 g kg -1 DM) and lignin (103-111 g kg -1 DM) content than cereal straws usually used in ruminant feeding, thus resulting in higher metabolizable energy (6.43-6.58 MJ kg -1 DM) content. In vitro digestibility was similar among lupin species (47.7-50.6%) and higher in pod shells (53.7%) than in straws (44.6%). Lupinus albus 'Estoril' and L. luteus 'Cardiga' presented considerable amounts of alkaloids in straws (23.9 and 119 mg kg -1 DM) and pod shells (20.5 and 298 mg kg -1 DM), while no alkaloids were detected in L. angustifolius 'Tango' biomass residues. Considering the combined production of straw and pod shells per lupin species, it is anticipated that lupin biomass residues produced per ha can fulfill 85% of the energy and nearly 50% of protein requirements of a flock of 4 to 9 dry and mid-pregnancy sheep with 50 kg body weight for one year. No negative effects on small (ovine) and large (bovine) ruminant species due to alkaloids are expected, even if biomass residues are consumed at up to 85% DM intake. The large production yield along with its nutritive value unveils the potential of lupin biomass residues valorization as alternative fodder for ruminants, promoting sustainability under a circular economy approach., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Maia, Monteiro, Valente, Sousa, Miranda, Castro, Cortez, Cabrita, Trindade and Fonseca.)

The stomach is a relevant spot of lipolysis for milk fat, but research on the effect of digested milk fat in the gastric epithelium is scarce and difficult to evaluate. In the present study, we implemented the semi-dynamic in vitro digestion model of INFOGEST, combined with gastric NCI-N87 cells, to study the effect of fat-free, whole conventional, and whole pasture-based milk on gastric epithelium. Cellular messenger ribonucleic acid (mRNA) expression of membrane fatty acids receptors (GPR41, GPR84), antioxidant enzymes (CAT, SOD, GPX), and inflammatory molecules (NF-κB p65, IL-1β, IL-6, IL-8 and TNF-α) was assessed. No significant differences were observed in mRNA expression of GPR41, GPR84, SOD, GPX, IL-6, IL-8, and TNF-α, after exposure of the NCI-N87 cells to milk digesta samples (p > 0.05). An increase of CAT mRNA expression was observed (p < 0.05), at a similar level, for all milk types. Whole milk digested samples induced higher mRNA expression of NF-κB p65 and IL-1β than fat-free milk (p < 0.05); while no differences were observed between whole conventional and whole pasture-based milk (p > 0.05). Moreover, the effect of milk digesta on gastric mRNA expression was studied in a scenario of subsequent stimulation of NCI-N87 monolayer with the pro-inflammatory cytokine IFN-γ. In these conditions, milk digesta samples increased CAT mRNA expression (p < 0.05), but had no effect in the expression of NF-κB p65 and IL-1β (p > 0.05). The increase of CAT mRNA expression suggests that milk fatty acids are used for energy production by gastric epithelial cells. Cellular antioxidant response to higher milk fatty acids availability could be associated to gastric epithelial inflammation, but did not contribute to increased inflammation in case of an external contact with IFN-γ. Besides, a conventional or a pasture-based origin did not affect the impact of whole milk in the NCI-N87 monolayer. The combined model responded to differences in milk fat content, which indicates its usefulness to study effects of foods at the gastric level., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Decomposition of dairy cows' excreta on housing floor leads to ammonia and greenhouse gases production, yet factors affecting total emissions have not been fully disclosed. This work aimed to assess the impact of lactation stage, feeding system and sampling time on gaseous emission potential of cow's faeces and urine in laboratory chambers systems. Individual faeces and urine were collected from two groups of four cows, at peak and post peak lactation, from three commercial farms with distinct feeding systems: total mixed ration (TMR), total mixed ration plus concentrate at robot (TMR + robot), and total mixed ration plus concentrate in automatic feeders (TMR + AF). Samples were collected before a.m. (T8h), at middle day (T12h), and before p.m. (T17h) milking. In a laboratory chambers system, faeces and urine were mixed in a ratio of 1.7:1, and ammonia and greenhouse gases emissions were monitored during 48-h. Cumulative N-N 2 O emissions were the highest in TMR + robot system, post peak cows and sampling time T17h. An interaction between stage of lactation and sampling time was detected for N-NH 3 and N-N 2 O (g/kg organic soluble N) emissions. Post peak cows also produced the highest cumulative N-NH 3 emissions. Overall results contribute for the identification of specific on-farm strategies to reduce gaseous emissions from cows' excreta., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

For the first time, seven European varieties of Vicia faba L. seeds were investigated for (1) their phytonutrients profile by HPLC-DAD-MS/MS, (2) total phenolic content (TPC), and (3) antioxidant capacity (DPPH and FRAP assays). A wide range of compounds were identified, namely alkaloids, organic acids, terpenoids, jasmonates, and phenolics; these two latter being the most abundant. TPC ranged between 2.62 and 4.3 mg (gallic acid equivalent) g -1 dry weight, for V. faba major variety Belshi and V. faba minor variety Bauska, respectively. The DPPH radical scavenging capacity showed poor correlation (r = 0.550, P = .041) with TPC, suggesting the presence of other antioxidant sources than phenolics. Still, FRAP was positively correlated with TPC (r = 0.709, P < .01) and DPPH (r = 0.819, P < .01). These results elucidated the phytonutrients and antioxidant properties of V. faba L. seeds as functional food sources., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The monitoring of the activity of alpha-amylase and amyloglucosidase is an important tool for studying their role in the hydrolysis of starch. Here we introduced an improved method capable to measure the activity of alpha-amylase and amyloglucosidase from different sources based on a quantitative starch-iodine assay. The developments of the assay sought the consistent preparation of the reagents, the rescale of the assay and the adjustment of the sensitivity. This was complemented by a glucose yield assay for amyloglucosidase that allowed a secondary source of information when insoluble starches were studied. The proposed method showed high precision in long-term use (RSD < 6.3%). Furthermore, all experimental conditions can be adapted according to the equipment available at each laboratory, transforming this method in a broadband analytical tool for screening alpha-amylase and amyloglucosidase activities. •Tailorable assay based on the starch-iodine staining for the determination of alpha-amylase and amyloglucosidase activities.•Enhanced consistence of reagent preparation.•High intra-day and inter-day reproducibility.

The present report studied the role of ultrasound (US) energy in the amyloglucosidase-based starch hydrolysis using two complementary approaches: (i) in the activity of six commercially-available amyloglucosidases (using soluble starch as substrate), and (ii) in the hydrolysis of four pure starches from different botanical sources. This corresponds to the first systematic evaluation of the role of US in starch hydrolysis mediated by amyloglucosidase, being a consequence of our previous report that assessed the effect of US in the activity of alpha-amylase (LWT - Food Science and Technology 84 (2017) 674-685). Regarding amlyloglucosidases, three enzymes obtained from Aspergillus niger (AN1-AN3), and Spirizyme Achieve (SPA), Spirizyme Fuel (SPF) and Spirizyme Ultra (SPU) were submitted to a Box-Behnken experimental design in order to establish the optimum conditions for their maximum activity. In the presence of US, we found both inactivation and activation, ranging from -88% (AN3) to 699% (SPA). The US promoted the enzyme activity when combined with lower temperatures (40-60 °C), with a marked effect in Spirizyme enzymes. Based on the optimum conditions established by the experimental design, we also evaluated the role of US in the glucose yield resulting from the hydrolysis of pure starches (corn, rice, potato, wheat). In this case, US led to higher glucose yields in all conditions tested. The enhancement factors observed ranged from 1.2 (AN1, rice starch) to 65 (SPA, potato starch) times. We compared these findings with previous reports, which highlighted the role of US in intensifying amyloglucosidase-based saccharification in mild conditions, by simultaneously influencing both enzyme and substrate. Hence, US power has to be fine-tuned for each particular enzyme in order to maximize process intensification., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Vicia faba L. pods are a by-product generated from the industrial processing of beans for human and animal consumption. As phenolic compounds may play important roles in health, the present work envisaged the phenolic characterization of seven European varieties and cultivars of V. faba (major and minor) pods and the assessment of their antioxidant activity. The V. faba methanolic extracts were characterized by HPLC-DAD-MS/MS for identification of polyphenolic compounds. The total phenolic content and antioxidant capacity of the extracts were evaluated by colorimetric methods (Folin-Ciocalteu, DPPH scavenging capacity assay, and FRAP assay). Main compounds identified by HPLC-DAD-MS/MS were derivatives of caffeic acid, coumaric acid and kaempferol. The broad bean Jögeva variety presented the highest content of free and esterified phenolics (26.3 and 26.7 mg 100 g -1 dry weight, respectively), followed by the horse bean varieties Bauska and Lielplatones. These results were corroborated by the analysis of total phenolic content, DPPH scavenging capacity and FRAP. This study confirmed the rich phenolic content of V. faba pods suggesting to be an interesting novel source for animal nutrition, promoting product quality and consumers' health., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This paper introduces a new method for a one-step determination of ammonia nitrogen (NH 3 ) in high complex solid and liquid samples from the agricultural and livestock sectors. To this end, we developed a simultaneous extraction and fluorimetric labeling of NH 3 , using gas diffusion microextraction (GDME), followed by the fluorescence measurement under 96-well microplate format. The GDME ensured a selective diffusion of NH 3 through a commercial hydrophobic membrane, and confined the acceptor solution, which included the fluorimetric labeling reagent o-phthalaldehyde (OPA). The OPA-NH 3 labeling reaction was optimized resorting to a full factorial experimental design, which showed that the reducing agent (Na 2 SO 3 ) concentration was critical to achieve the highest sensitivity. A similar optimization approach for GDME showed that time and temperature significantly influenced the sensitivity of the assay, and also that the modifications in these two factors could be used to adjust the sensitivity according to the concentrations present in the samples. In our final conditions, it was possible to quantify NH 3 in the range between 0.38 and 6.27mgL -1 using a 10min extraction at 25°C in different samples (e.g., corn and grass silages, feces, urine). The developed method showed a high repeatability and reproducibility (intraday and interday relative standard deviations values of 4.5% and 9.5%, respectively) and an adequate limit of detection (0.22mgL -1 ). This new methodology also highlighted the simplicity and versatility of GDME for the determination of volatile components of high-complex matrices, which will certainly drive future developments in the analysis of environmental and biological samples., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Twenty-nine mature raw varieties of grain legume seeds (chickpeas, field peas, faba beans, common vetch and lupins) produced in Europe were investigated for their phenolic profile by means of high performance liquid chromatography coupled to diode array detection (HPLC-DAD). To the best of our knowledge, this study reported for the first time the phenolic composition of mature raw seeds of chickpea type Desi, field pea and common vetch. Phenolic acids were predominant compounds in chickpeas, field peas and common vetch compared to flavonoids, whereas the opposite was observed for lupin seeds. Yellow lupins presented the highest levels of total phenolic compounds followed by narrow-leafed lupins (in average 960 and 679mg/kg, dry basis, respectively), whereas Kabuli chickpeas got the lowest ones (in average 47mg/kg, dry basis). Principal component analysis revealed that flavones and total levels of phenolic compounds were responsible for nearly 51% of total data variability., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The fatty acid (FA) analysis of grass and maize silages was studied by application of a direct transesterification method (DT) followed by purification by solid-phase extraction (SPE). The choice of the internal standard (IS) for quantification of FA by gas-liquid chromatography (GLC) was also studied. The acidic DT method applied to grass silage samples produced a high amount of non-fatty acid methyl ester compounds (non-FAME) compared with those formed in maize silages. The application of the SPE cleanup step reduced significantly the amount of non-FAME compounds in both samples. Five FAs were tested as IS; among them, 3 were naturally present in all silages, however their use as IS did not affect quantification of total FA composition. Nevertheless, some minor FAs present in silages were significantly affected by the IS used. Additionally, application of corrections to the GLC peak areas did not significantly influence quantification of total FA composition of silages.

Direct transesterification (DT) and solvent extraction with acid or basic derivatization procedure for fatty acid (FA) analysis in herbage were compared. The highest total FA, alpha-linolenic and linoleic acid contents were obtained with DT. However, DT also produced the highest amount of interfering compounds, identified as phytadienes and sugar derivative products, which may co-elute with FA. An additional step based on solid-phase extraction to produce clean samples was proposed. This procedure is simple and gives good recoveries for the FA fortified samples. Additionally, structural characterization of 16:1 trans-3 was conducted by covalent adduct chemical ionization tandem mass spectrometry.