Production of dairy cows fed whole-crop cereals or ryegrass silages supplemented with a fixed amount of concentrate

Abstract

Feed intake and milk production responses to whole-crop cereal silages and ryegrass silage (RGS) supplemented with a fixed amount of concentrate were measured in three 4×4 Latin square-designed experiment with three 17-day periods using 12 Holstein cows. Diets consisted of a fixed amount of concentrate (10 kg) and one of the following silages offered ad libitum: RGS, Mondego wheat silage (MWS), Alva wheat silage (AWS), and triticale silage (TS). Silage dry matter intake and milk yield were significantly higher for RGS. Milk composition was not affected by silage treatment. The whole-crop cereal silages tended to show higher milk concentrations of anteiso C15:0, iso C15:0, C16:1 and C18:1 and lower concentrations of C18:2 and C18:3. This study suggests better response of dairy cows to a single cut RGS than to whole-crop cereal silages harvested in an early stage of maturity.     

A new lupene triterpenetriol and anticholinesterase activity of Salvia sclareoides

A new lupene triterpenetriol was isolated from the acetone extract of the aerial parts of Salvia sclareoides and characterised as (1beta,3beta)-lup-20(29)-ene-1,3,30-triol (1). In addition, nepetidin (2), nepeticin (3), lupendiol (4), (1beta,11alpha)-dihydroxy-lup-20(29)-en-3-one (5), ursolic acid (6), sumaresinolic acid (7) and hederagenin (8), were identified in this Salvia sp. To the best of our knowledge, the compounds 2 and 7 are new constituents in Salvia spp. The acetone, ethanol, butanol and water extracts of the plant were screened for the in vitro inhibitory activity of acetylcholinesterase (AChE) and butyrilcholinesterase (BChE), enzymes which play a role in the Alzheimer disease. All extracts inhibited acetylcholinesterase activity at 10 microg/ml, a remarkable activity since the standard drug rivastigmine does not inhibit acetylcholinesterase at the same concentration. Regarding the butyrilcholinesterase, the acetone extract at 1000 microg/ml was able to inhibit completely the enzyme activity and the butanol and ethanol extracts, at this concentration, produced a potent inhibition of BchE.     

A novel pentacyclic triterpene from Leontodon filii

A novel oleanene triterpenetetrol was isolated from the chloroform extract of the aerial parts of Leontodon filii. Its structure was shown to be 2beta,3beta,15alpha,21beta-olean-12-ene-2,3,15,21-tetrol by chemical and spectroscopic methods. The fungicidal efficacy of the chloroform and methanol extracts of the plant was also evaluated, a protective effect being found against Plasmopara viticola, Botrytis cinerea, particularly powerful against Pyricularia oryzae.     

The prion-like protein Doppel enhances ovine spermatozoa fertilizing ability

The function of prion‐like protein Doppel was suggested to be related to male fertility. In this study, the importance of ovine Doppel polypeptide on spermatozoa capacitation and fertilization was evaluated. After refolding, recombinant Doppel (rDpl) was supplemented with different concentrations (40, 80 or 190 ng/ml) to ovine spermatozoa during the capacitation process. In experiment 1, post‐thawed ovine spermatozoa were incubated with different concentrations of rDpl during 1 h for swim‐up, and changes in sperm motility, concentration, vigour, viability and capacitation were monitored (10 replicates). In experiment 2, the fertilization ability of post‐swim‐up spermatozoa incubated as above was tested through heterologous fertilization of bovine in vitro matured oocytes (n = 423, three replicates). Regardless of dosage, rDpl improved (p ≤ 0.03) spermatozoa viability. Sperm individual motility and vigour were the highest (p ≤ 0.04) for the group receiving 190 ng/ml rDpl. Sperm supplemented with the highest doses of rDpl achieved higher (p = 0.02) fertilization rates (56.0 ± 3.0%) than control (39.1 ± 2.2%) and 40 ng/ml rDpl (39.8 ± 2.7%). Preliminary data suggest that Doppel protein may enhance in vitro spermatozoa fertilizing ability.     

Assessment of genetic relationships among Pyrus species and cultivars using AFLP and RAPD markers

Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.     

Effect of ensiling and silage additives on fatty acid composition of ryegrass and corn experimental silages

Two experiments were conducted using laboratory mini-silos to study the effect of ensiling and silage additives on fatty acid (FA) composition, including minor or unusual FA, of ryegrass and corn silages. Ryegrass was ensiled for 12 wk with no additives, with the addition of a bacterial inoculant or formic acid. Corn was ensiled for 9 wk without additives, with the addition of a bacterial inoculant or calcium formate. Ensiling affected both total FA content and FA composition of ryegrass silages. Total FA concentration increased (P < 0.001) during ryegrass ensiling. The proportions (g/100 g of total FA) of the major unsaturated FA, 18:3n-3 and 18:2n-6, were not affected (P > 0.05) by ensiling. However, their concentration (mg/g of DM) in silages was greater (P=0.017 and P=0.001, respectively) than in fresh ryegrass. Two 18:2 FA (trans-11,cis-15 and cis-9,cis-15) that were not originally present in the fresh ryegrass were detected in silages. Silage additives affected the FA composition of ryegrass silages, mostly by increasing the proportions of SFA, but not on total FA concentration. Ensiling did not affect (P=0.83) total FA content of corn silages; however, FA composition was affected, mostly by decreasing the proportions of 18:2n-6 and 18:3n-3. Silage additives had no effect on corn silage FA composition. Exposing corn silages to air resulted in no oxidation of FA or reduction in total FA content or composition.     

Structural, physical, and chemical modifications induced by microwave heating on native agar-like galactans

Native agars from Gracilaria vermiculophylla produced in sustainable aquaculture systems (IMTA) were extracted under conventional (TWE) and microwave (MAE) heating. The optimal extracts from both processes were compared in terms of their properties. The agars' structure was further investigated through Fourier transform infrared and NMR spectroscopy. Both samples showed a regular structure with an identical backbone, β-d-galactose (G) and 3,6-anhydro-α-l-galactose (LA) units; a considerable degree of methylation was found at C6 of the G units and, to a lesser extent, at C2 of the LA residues. The methylation degree in the G units was lower for MAE(opt) agar; the sulfate content was also reduced. MAE led to higher agar recoveries with drastic extraction time and solvent volume reductions. Two times lower values of [η] and M(v) obtained for the MAE(opt) sample indicate substantial depolymerization of the polysaccharide backbone; this was reflected in its gelling properties; yet it was clearly appropriate for commercial application in soft-texture food products.     

Electric ultrafreezer (− 150 °C) as an alternative for zebrafish sperm cryopreservation and storage

Zebrafish sperm cryopreservation is a fundamental methodology to manage and back-up valuable genetic resources like transgenic and mutant strains. Cryopreservation usually requires liquid nitrogen for storage, which is expensive and hazardous. Our objective was to evaluate if electric ultrafreezers (??150 °C) are a viable alternative for zebrafish sperm storage. Zebrafish sperm was cryopreserved in the same conditions (??20 °C/min), stored either in liquid nitrogen or in an ultrafreezer, and thawed after 1 week, 1 month, and 3 months. Sperm motility, membrane integrity, and fertilization ability were assessed. There were no significant differences in motility and hatching rate throughout storage time. Additionally, we aimed at understanding if cryopreservation directly in an ultrafreezer (??66 °C/min) could improve post-thaw sperm quality. Freezing at ??20 °C/min was performed as before, and compared to samples cryopreserved with a fast cooling rate by placing directly in an ultrafreezer (??66 °C/min). Sperm quality was assessed according to motility, viability, DNA fragmentation, and apoptosis (annexin V). The ??66 °C/min cooling rate showed significantly higher membrane and DNA integrity, and lower number of cells in late apoptosis in comparison to the other treatments. This study showed that zebrafish sperm cryopreservation and storage in an ultrafreezer system is possible and a fast cooling rate directly in ultrafreezer improves post-thaw sperm quality.     

Technical note: stearidonic acid metabolism by mixed ruminal microorganisms in vitro

Dietary supplementation of stearidonic acid (SDA; 18:4n-3) has been considered a possible strategy to increase n-3 unsaturated fatty acid content in ruminant products; however, little is known about its metabolism in the rumen. In vitro batch incubations were carried out with bovine ruminal digesta to investigate the metabolism of SDA and its biohydrogenation products. Incubation mixtures (4.5 mL) that contained 0 (control), 0.25, 0.50, 0.75, 1.00, 1.25, or 1.50 mg of SDA supplemented to 33 mg (DM basis) of commercial total mixed ration based on corn silage, for dairy cows, were incubated for 72 h at 39°C. The content of most fatty acids in whole freeze-dried cultures was affected by SDA supplementation. Branched-chain fatty acids decreased linearly (P < 0.01), and odd-chain fatty acids decreased quadratically (P < 0.01), particularly from 1.00 mg of SDA and above, whereas most C18 fatty acids increased linearly or quadratically (P ≤ 0.04). Stearidonic acid concentrations at 72 h of incubation were very small (<0.6% of total fatty acids and ≤0.9% of added SDA) in all treatments. The apparent biohydrogenation of SDA was extensive, but it was not affected by SDA concentration (P > 0.05). Biohydrogenation followed a pattern similar to that of other C18 unsaturated fatty acids up to 1.00 mg of SDA. Stearic acid (18:0) and vaccenic acid (18:1 trans-11) were the major fatty acids formed, with the latter increasing 9-fold in the 1.00 mg of SDA treatment. At greater inclusion rates, 18:0 and 18:1 trans isomers decreased (P ≤ 0.03), accompanied by increases in unidentified 18:3 and 18:4 isomers (P = 0.02), suggesting that the biohydrogenation pathway was inhibited. The present results clearly indicate that SDA was metabolized extensively, with numerous 18:4 and 18:3 products formed en route to further conversion to 18:2, 18:1 isomers, and 18:0.     

Anti-Helicobacter pylori activity of Terminalia macroptera root

The root of Terminalia macroptera Guill. & Perr. (Combretaceae) is widely used in African traditional medicine to treat various infectious diseases, including stomach-associated diseases. This study investigates the in vitro activity of T. macroptera root extract against reference strains and clinical isolates of H. pylori and attempts to localize the extract bioactivity. T. macroptera hydroethanol (80% V/V) root extract (Tmr) activity was tested against three standard strains and sixty two clinical strains of H. pylori. Tmr liquid-liquid partition fractions were screened against twenty H. pylori strains. Qualitative analysis of Tmr and its fractions was performed by HPLC-UV/DAD. The antibiotic characterization of the H. pylori strains revealed that 20% of the tested clinical isolates were resistant to at least two of the three antibiotics belonging to the main groups of antibiotics used in multi-therapy to eradicate H. pylori infections. In contrast, Tmr showed anti-H. pylori activity against the majority (92%) of the tested strains (MIC(50) and MIC(90)=200 μg/ml). The Tmr water liquid-liquid fraction (Tmr-3) and the precipitate obtained from this fraction (Tmr-5) were the most active tested samples, showing a MIC(50) of 100 μg/ml. The present work proves the in vitro activity of T. macroptera against H. pylori, thus confirming the utility of this traditional medicinal plant to treat stomach complaints due to H. pylori infection. The main compounds of Tmr and of Tmr-3 were the ellagitannins terchebulin and punicalagin. These compounds can be considered as markers of T. macroptera root active extracts against H. pylori.