In vitro ruminal biohydrogenation of eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic acid (DHA) in cows and ewes: Intermediate metabolites and pathways

A great deal of uncertainty still exists about intermediate metabolites and pathways explaining the biohydrogenation (BH) of 20- and 22-carbon polyunsaturated fatty acids (PUFA). Therefore, this study was conducted to provide further insight into the ruminal metabolism of 20:5 n-3 (EPA), 22:5 n-3 (DPA), and 22:6 n-3 (DHA), the main n-3 PUFA present in the marine lipids used in dairy ruminant feeding, and to examine potential differences between bovine and ovine. To meet this aim, we investigated the 20- and 22-carbon metabolites accumulated during in vitro incubation of EPA, DPA, and DHA with rumen inocula from cows and ewes. The PUFA were added at a dose of 2% incubated dry matter and digesta samples were analyzed after 24 h of incubation using complementary gas-liquid chromatography of fatty acid methyl esters and gas chromatography-mass spectrometry of 4,4-dimethyloxazoline derivatives. Results suggested that the main BH pathway of EPA and DPA would proceed via the reduction of the double bond closest to the carboxyl group (cis-5 in EPA and cis-7 in DPA); curiously, this mechanism seemed of much lower importance for DHA. Thus, DPA would not be a major intermediate product of DHA and their BH might actually follow separate pathways, with the accumulation of numerous unique metabolites in each case. A principal component analysis supported this hypothesis, with a clear separation between PUFA treatments in the score and loading plots. Within EPA and DPA groups, cow and ewe samples loaded separately from each other but not distant. No conjugated 20:5, 22:5, or 22:6 isomer compatible with the initial product of EPA, DPA, or DHA metabolism, respectively, was identified in the ruminal digesta, although this would not unequivocally exclude their transient formation. In this regard, results from DPA incubations provided the first indication that the metabolism of this very long chain PUFA may involve the formation of conjugated double bond structures. The BH of EPA, DPA, and DHA resulted in the appearance of several tentative trans-10–containing metabolites, showing a general trend to be more abundant in the digesta of ewes than in that of cows. This finding was speculated to have some relationship with the susceptibility of dairy sheep to marine lipid-induced milk fat depression. Differences in the relative proportion of intermediate products would also suggest an influence of ruminant species on BH kinetics, with a process that would likely be slower and less complete in cows than in ewes.     

Rapid physicochemical characterization of innovative fucoidan/fructan powders by ATR–FTIR

Functional food has been highly demanded lately because of its benefits in counteracting diseases. Fucoidan and agave fructan are ingredients that enhance the growth of beneficial bacteria in the gut (prebiotics). This mixture has great potential to develop innovative products but it has never been explored before. Because of fucoidan is more expensive than agave fructan, the innovative proposed mixture is vulnerable to adulteration. This research was aimed to assess the accuracy of Fourier transform infrared spectroscopy with attenuated total reflectance (ATR–FTIR) coupled with chemometrics to identify and predict concentration of both polysaccharides in powder mixtures (0–100%). Absorption bands at 1240–1255 and 836–840 cm^?1 were attributed to fucoidan and a strong peak at ~ 936 cm^?1 confirmed the fructan presence. Peak areas were best fitted into linear models (\({\text{R}}_{\text{adj}}^{2}\) ≥ 0.92, RMSE ≤ 3.54%). This achievement may be useful to certificate ingredients contained in fucoidan–fructan mixtures, preventing adulteration.     

Optimization of ultrasonic-assisted extraction of phenolic compounds from <Emphasis Type="Italic">Justicia spicigera</Emphasis> leaves

A Box–Behnken design (Extraction-time, pulse-cycle, sonication-amplitude) was employed to extract phenolic compounds from Justicia spicigera leaves by ultrasonic-assisted extraction. The muicle leaves extracts were analyzed measuring total phenolic compounds and antioxidant capacity. According to response surface methodology the optimal conditions of ultrasonic-assisted extraction to obtain the highest soluble phenolic content were 2 min (extraction time) for 0.7 s (pulse cycle) at 55% of sonication amplitude. Under these optimal conditions, the total phenolic content was higher when was used ultrasonic-assisted extraction (54.02 mg/g) than stirring (46.46 mg/g) and thermal decoction (47.76 mg/g); however, the antioxidant capacity from J. spicigera extracts did not increase by ultrasonic-assisted extraction. The extracts or aqueous infusions from J. spicigera leaves are used for therapeutic proposes, therefore the ultrasonic-assisted extraction is a useful technology to improve the extraction of phytochemicals from J. spicigera leaves.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).     

Altering the ratio of dietary palmitic,stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility,energy partitioning,and production responses of dairy cows

The objective of this study was to evaluate the effects of varying the ratio of dietary palmitic (C16:0), stearic (C18:0), and oleic (cis-9 C18:1) acids in basal diets containing soyhulls or whole cottonseed on nutrient digestibility, energy partitioning, and production response of lactating dairy cows. Twenty-four mid-lactation multiparous Holstein cows were used in a split-plot Latin square design. Cows were allocated to a main plot receiving either a basal diet with soyhulls (SH, n = 12) or a basal diet with whole cottonseed (CS, n = 12) that was fed throughout the experiment. Within each plot a 4 × 4 Latin square arrangement of treatments was used in 4 consecutive 21-d periods. Treatments were (1) control (CON; no supplemental fat), (2) high C16:0 supplement [PA; fatty acid (FA) supplement blend provided ～80% C16:0], (3) C16:0 and C18:0 supplement (PA+SA; FA supplement blend provided ～40% C16:0 + ～40% C18:0), and (4) C16:0 and cis-9 C18:1 supplement (PA+OA; FA supplement blend provided ～45% C16:0 + ～35% cis-9 C18:1). Interactions between basal diets and FA treatments were observed for dry matter intake (DMI) and milk yield. Among the SH diets, PA and PA+SA increased DMI compared with CON and PA+OA treatments, whereas in the CS diets PA+OA decreased DMI compared with CON. The PA, PA+SA, and PA+OA treatments increased milk yield compared with CON in the SH diets. The CS diets increased milk fat yield compared with the SH diets due to the greater yield of de novo and preformed milk FA. The PA treatment increased milk fat yield compared with CON, PA+SA, and PA+OA due to the greater yield of mixed-source (16-carbon) milk FA. The PA treatment increased 3.5% fat-corrected milk compared with CON and tended to increase it compared with PA+SA and PA+OA. The CS diets increased body weight (BW) change compared with the SH diets. Additionally, PA+OA tended to increase BW change compared with CON and PA and increased it in comparison with PA+SA. The PA and PA+OA treatments increased dry matter and neutral detergent fiber digestibility compared with PA+SA and tended to increase them compared with CON. The PA+SA treatment reduced 16-carbon, 18-carbon, and total FA digestibility compared with the other treatments. The CS diets increased energy partitioning toward body reserves compared with the SH diets. The PA treatment increased energy partitioning toward milk compared with CON and PA+OA and tended to increase it compared with PA+SA. In contrast, PA+OA increased energy partitioned to body reserves compared with PA and PA+SA and tended to increase it compared with CON. In conclusion, milk yield responses to different combinations of FA were affected by the addition of whole cottonseed in the diet. Among the combinations of C16:0, C18:0, and cis-9 C18:1 evaluated, fat supplements with more C16:0 increased energy output in milk, whereas fat supplements with more cis-9 C18:1 increased energy storage in BW. The combination of C16:0 and C18:0 reduced nutrient digestibility, which most likely explains the lower performance observed compared with other treatments.     

Identification of components of Brazilian honey by 1H NMR and classification of its botanical origin by chemometric methods

The potential of NMR spectroscopy to differentiate honeys concerning to the nectar employed in its production was evaluated. The application of chemometric methods to ¹H NMR spectra has allowed to discriminate the honeys produced in the state of São Paulo, being identified the signals of responsible substances for the discrimination. Application of PCA and HCA methods to ¹H NMR data have resulted in the natural clustering of the samples. Wildflower honeys were characterized by higher concentration of phenylalanine and tyrosine. Citrus honeys showed higher amounts of sucrose than other compounds, while eucalyptus honeys had higher amount of lactic acid than the others. Assa-peixe honeys showed spectra similar to eucalyptus and citrus. Sugar-cane honeys showed some signals similar to eucalyptus and citrus honeys, but also showed the tyrosine and phenylalanine signals. Adulterated honeys showed 5-hydroxymethylfurfural, citric acid and ethanol signals. KNN, SIMCA and PLS-DA methods were used to build predictive models for honey classification. In the commercial honeys prediction KNN, SIMCA and PLS-DA models correctly classified 66.7; 22.2 and 72.2% of the samples, respectively.