Dario Colombatto

A protease additive increases fermentation of alfalfa diets by mixed ruminal microorganisms in vitro.

In vitro experiments were conducted to examine the characteristics and mode of action of a protease that increased the ruminal fiber digestibility of alfalfa hay. A commercial source of protease (Protex 6L, Genencor Int., Rochester, NY), already characterized for its main activities, was further analyzed to determine protease activity in response to pH, molecular size by SDS-PAGE, specificity to degrade model or feed substrates, response to autoclaving, and action of specific protease inhibitors in the absence or presence of ruminal fluid. In addition, batch culture in vitro incubations in buffered ruminal fluid were conducted to compare the enzyme product with purified protease sources, and dose-response studies (0 to 10 microL/g of forage DM) were carried out using alfalfa hay as a substrate. The enzyme product was shown to be an alkaline protease (optimum pH >8.5) of approximately 30 kDa. Specificity in the absence of ruminal fluid showed that the enzyme was active against gelatin and casein to the same extent, whereas it had limited (21% of the total) activity on BSA. In the presence of ruminal fluid and with the use of feed substrates, the protease increased (P < 0.05) 22-h IVDMD (%) of alfalfa hay, fresh corn silage, dry-rolled corn, and a total mixed ration composed of the 3 ingredients (39.5 vs. 44.7; 50.3 vs. 54.5; 63.8 vs. 68.4; and 55.4 vs. 56.4 for control vs. protease for each feed, respectively). Inhibitor studies in the absence of ruminal fluid indicated that the enzyme was inhibited most by a serine protease inhibitor but not by cysteine- or metalloprotease inhibitors (10 vs. 1.9 and 0.1%, respectively). In the presence of ruminal fluid, the serine protease inhibitor reversed (P < 0.05) the increase in alfalfa IVDMD achieved by the enzyme product, such that IVDMD was similar to that of the control treatment. Comparisons among different proteases revealed that only pure subtilisin achieved increases in IVDMD that were similar to those with protease, suggesting the serine protease was subtilisin-like (EC 3.4.1.62). Dose-response studies using alfalfa hay as substrate showed quadratic responses in IVDMD, NDF digestion, and hemicellulose and protein disappearance. It is postulated that this enzyme acts by removing structural proteins in the cell wall, allowing ruminal microbes to gain faster access to digestible substrates.

Nutritional and eating quality of Argentinean beef : A review

This review deals with distinctive aspects of quality of Argentinean beef in terms of tenderness, flavour, colour, juiciness, taste, acceptability, lipid content and composition and its resultant nutraceutical characteristics. Differences are due to beef production systems based on temperate or tropical grasslands aimed at shortening the fattening phase as far as possible, with limited or null use of concentrates. However, the effect of limited supplemental feeding is also discussed as well as the responses arising from the use of beef cattle genotypes, including British, Continental, Dairy, Zebu breeds and their crosses, adapted to the various environments and systems found in the country.

Essential oils from Lippia turbinata and Tagetes minuta persistently reduce in vitro ruminal methane production in a continuous-culture system

The aim of the present study was to evaluate the impact of essential oils (EO) from Lippia turbinata (LT) and Tagetes minuta (TM) as well as the rotation of both EO on fermentation parameters in vitro. Daily addition of LT, TM, or a 3-day rotation between them (TM/LT), as well as a control (without EO), was evaluated using the rumen simulation technique (Rusitec). The experiment lasted 19 days, with a 7-day adaptation period, followed by 12 days of treatment (Days 0–12). The EO were dissolved in ethanol (70% vol/vol) to be added daily to fermenters (300 μL/L) from Day 0. Daily measurements included methane concentration, total gas production, apparent DM disappearance and pH, which started 2 days before the addition of treatments. On Days 0, 4, 8 and 12 apparent crude protein disappearance and neutral detergent fibre disappearance, ammonia and volatile fatty acid concentration and composition were determined. Methane production was significantly inhibited shortly after addition of both EO added individually, and persisted over time with no apparent adaptation to EO addition. The TM/LT treatment showed a similar effect on methane production, suggesting that rotating the EO did not bring further improvements in reduction or persistency compared with the inclusion of the EO individually. Gas production, total volatile fatty acid concentration and composition and apparent crude protein disappearance were not affected by EO addition. Compared with the control, a 5% reduction of apparent DM disappearance and a 15% reduction of neutral detergent fibre disappearance were observed with the addition of EO. Only TM and TM/LT reduced ammonia concentration. Given the significant and persistent antimethanogenic activity of both EO, and the potential of T. minuta to modify nitrogen metabolism, EO from these plant species are of interest for developing new feed additives with potential application in ruminant nutrition that are also likely to be acceptable to consumers.

Anatomy, nutritional value and cell wall chemical analysis of foliage leaves of Guadua chacoensis (Poaceae, Bambusoideae, Bambuseae), a promising source of forage.

BACKGROUND The present study combines morphological and anatomical studies, cell wall chemical composition analysis, as well as assessment of the nutritional value of Guadua chacoensis foliage leaves. RESULTS Foliage leaves of G. chacoensis are a promising source of forage because: (a) as a native woody bamboo, it is adapted to and helps maintain environmental conditions in America; (b) leaf anatomical studies exhibit discontinuous sclerenchyma, scarcely developed, while pilose indumentum, silica cells, prickles and hooks are also scarce; (c) it has a high protein content, similar to that of Medicago sativa, while other nutritional parameters are similar to those of common forages; and (d) glucuronoarabinoxylan, the major extracted polysaccharide, has one-third of the 4-linked β-d-xylopyranosyl units of the backbone substituted mainly with α-l-arabinofuranose as single stubs or non-reducing end of short chains, but also 5-linked α-l-arabinofuranose units, terminal β-d-xylopyranose and d-galactopyranose units, as well as α-d-glucuronic acid residues and small amounts of its 4-O-methylated derivative. CONCLUSION These results constitute the first report on this species, and as culms are utilized in constructions and crafts, the remaining leaves, when used as forage, constitute a byproduct that allows an additional income opportunity.