Effect of zinc supplementation from two sources on growth,nutrient utilization and immune response in male crossbred cattle (Bos indicus×Bos taurus) bulls

To investigate effects of Zn supplementation on performance, mineral balance and immune response, 15 male crossbred cattle (Bos indicus×Bos taurus) bulls of about 14 ± 0.4 months of age and weighing 226.0 ± 9.1 kg were divided in to three groups of five. Bulls in the control group were fed wheat straw and a concentrate mixture (basal diet with 32.5 mg Zn/kg dry matter (DM)), and in ZnSO₄ and ZnProp groups 35 mg Zn/kg DM was added through Zn sulphate and Zn propionate, respectively. All bulls were fed their respective treatment diets for 180 days. Daily feed intake was recorded and bulls were weighed at every 15 days to determine change in body weight (BW). After 120 days of feeding, bulls were vaccinated with Brucella abortus strain 19, and cell mediated and humoral immune responses were assessed between 120 and 148 days of experimental feeding. After 150 days of feeding, a metabolism study of 6 days duration was completed to determine nutrient digestibility and mineral balances (i.e., Ca, P, Zn, Cu, Fe and Mn). Intake of total DM, digestibility of DM, crude protein, ether extract, neutral detergent fibre and acid detergent fibre, N balance, average daily gain, feed: gain did not differ between the groups. Intake, excretion and balance of Ca, P, Zn, Cu, Fe and Mn also did not differ between the groups. However, retention of Zn was higher (P<0.001) in the ZnProp group. Bulls supplemented with Zn propionate had higher cell mediated (P<0.01) and humoral (P<0.05) immune response, while there was no alteration in immune response due to Zn sulphate supplementation. Results indicate that a diet containing about 32.5 mg Zn/kg DM was adequate to support normal growth and digestibility, but a better immune response occurred when Zn propionate was added to the diet at 35 mg/kg DM versus Zn sulphate.     

In vitro ruminal methanogenesis of a hay-rich substrate in response to different combination supplements of nitrocompounds; pyromellitic diimide and 2-bromoethanesulphonate

Sixteen combinations of 5 treatments at 4 levels were designed in a L₁₆(4⁵) orthogonal experimental design to evaluate associative effects of five methanogenesis inhibitors at four dose levels: nitroethane (NE, 0 mM, 5 mM, 10 mM and 15 mM), 2-nitroethanol (NEOH, 0 mM, 5 mM, 10 mM and 15 mM), 2-nitro-1-propanol (NPOH, 0 mM, 5 mM, 10 mM and 15 mM), pyromellitic diimide (PMDI, 0 mM, 0.02 mM, 0.05 mM and 0.07 mM) and 2-bromoethanesulphonate (BES, 0 mM, 0.01 mM, 0.03 mM and 0.05 mM) on in vitro ruminal methane production of the mixed substrate (Chinese wildrye hay:maize meal = 4:1) using a cumulative gas production technique. After 48 h incubation, in vitro dry matter disappearance (IVDMD), total gas production (GP₄₈, ml/g DM) and total volatile fatty acids (VFA) production in various combinations of these inhibitors were decreased by 10.6-56.0, 26.5-44.5 and 20.3-47.6%, respectively (P<0.05). The molar proportion of acetate in the inhibitor combination groups was decreased by 6.6-12.5% while those of propionate and butyrate were increased by 7.0-19.2 and 21.9-56.5% (P<0.01), respectively. Methane proportion (MP) in total gas production was reduced by 79.4-98.5% (P<0.01), and the highest inhibition occurred in the combination of 10 mM NE, 10 mM NPOH, 0.07 mM PMDI and 0.01 mM BES in cultures. The partial correlation coefficients between NE, NEOH, NPOH, PMDI or BES and CH₄ proportion were −0.465 (P<0.01), −0.417 (P<0.01), −0.355 (P<0.05), −0.408 (P<0.01) and −0.345 (P<0.05), respectively, indicating that NE was the most potent inhibitor, followed by NEOH and PMDI, and finally NPOH and BES. In general, VFA production in the inhibitor combinations was substantially shifted to produce much more butyrate and propionate and less acetate. The combination of 15 mM NE, 10 mM NEOH, 5 mM NPOH, 0.07 mM PMDI and 0.01 mM BES in cultures, leading to >95% methane inhibition, may be the optimal application of these inhibitors with less depression of total VFA production. Further feeding trials to validate these combinations is still required on rumen function, methane production, growth performance and milk production.     

Influence of method of whole-wheat feeding on the performance,digestive tract development and carcass traits of broiler chickens

The present study was conducted to investigate the effects of feeding whole wheat either through a mixed feeding (MF) or free choice feeding (FCF) system on the performance, digestive tract development and carcass traits of broiler chickens. The following three treatments, based on wheat and soybean meal, were employed: GW, ground-wheat diet with 600–690 g wheat kg⁻¹; MF, GW diet with 490–500 g wheat kg⁻¹ and 100–200 g whole wheat kg⁻¹; and FCF, whole wheat and a protein concentrate offered in separate feeders. Each diet was fed to six pens of 36 birds each from day 7 to 35 post-hatch. Over the 7–35-day trial period, no differences (P>0.05) were observed between the weight gain, feed intake and feed per gain of broilers receiving the GW and MF treatments. Birds receiving the FCF treatment had the lowest (P<0.05) weight gain and feed intake, and the highest (P<0.05) feed per gain. During week 1 of the trial, the protein concentrate was consumed more than the whole wheat (0.69 vs. 0.31) in the FCF treatment, resulting in the amount of concentrate offered being restricted during subsequent weeks. Over the trial period, the average consumption of protein concentrate remained high (0.56 of the total intake). Both whole-wheat treatments increased (P<0.05) the relative gizzard weights. Factors that affect diet selection such as learning and previous experience, visual differences between the foods, texture and flavour of the food, and palatability may explain the lower whole-wheat intake in the FCF treatment. In this study, whole wheat was introduced only on day 7 and it is possible that the birds may have to be trained to experience choice feeding from the first week of life. However, the present results suggest that FCF may not be an appropriate feeding system for fast growing modern broilers.     

Nutritional characterization of Mucuna pruiriens: 2. In vitro ruminal fluid fermentability of Mucuna pruriens,Mucunal-dopa and soybean meal incubated with or without l-dopa

Three in vitro experiments were conducted to determine the rumen fermentability of Mucuna (M) pruriens (24 g 3,4-dihydroxy-l-phenylalanine (l-dopa)/kg dry matter (DM) and soybean meal treated with (SBD) or without (SB) 138 g l-dopa/kg DM). Additional objectives were to determine if l-dopa inhibits rumen fermentation, and if ruminal microbes can adapt to l-dopa or M. In Experiment 1, ground (1 mm) substrates were incubated in triplicate at 38 °C in 9 ml nutrient media and 1 ml rumen fluid in a series of six, 48 h, consecutive batch cultures. The first culture was inoculated with rumen fluid from two donor cows. Subsequent cultures were inoculated with fluid (1 ml) from the previous culture. The DM digestibility (DMD, 616 g/kg vs. 540 g/kg; P<0.01) and gas production (51.7 ml/g vs. 44.2 ml/g DM; P<0.05) were higher from fermentation of M versus SB but similar for SB and SBD (540 g/kg vs. 554 g/kg and 44.2 ml/g DM vs. 43.5 ml/g DM, respectively). The slopes of the relationships between DMD (g/kg) or gas production (ml/g DM) and fermentation period were not reduced by fermenting M (−0.014 DMD slope; 2.28 gas production slope) or SBD (−0.014 DMD slope; 0.459 gas production slope), instead of SB (−0.002 DMD slope; 1.039 gas production slope), indicating microbial adaptation to M and SBD. Total volatile fatty acid concentration (VFA; 53.7, 54.9 and 54.9 mmol/l) and molar proportions of VFA were similar among substrates. Gas production kinetics of M versus SB (Experiment 2), and SB versus SBD (Experiment 3) were also measured after substrates were incubated in triplicate in buffered rumen fluid for 24 h using a non-linear exponential model to fit the data. Residual l-dopa was measured after separate fermentation of substrates in triplicate for 0, 4, 8, 16 and 24 h. Fermentation of M versus SB produced more (P<0.05) gas (250 ml/g vs. 100 ml/g DM) and total VFA (203 mmol/l vs. 180 mmol/l) and a lower (P<0.05) acetate:propionate ratio (1.35 vs. 1.87; P<0.05). Adding l-dopa to SB increased (P<0.01) gas production (92 ml/g DM vs. 200 ml/g DM), and total VFA concentration (132 mmol/l vs. 188 mmol/l), but reduced (P<0.05) gas production rate (0.08 ml/h vs. 0.05 ml/h). The concentration of l-dopa in fermented M and SBD decreased by 53 and 47%, respectively during fermentation. In conclusion, M was more fermented than SB and degradation of l-dopa during ruminal fermentation and microbial adaptation to l-dopa were confirmed. Adding l-dopa to SB did not impair ruminal fermentation.     

Effects of exogenous enzymes on in vitro gas production kinetics and ruminal fermentation of four fibrous feeds

This study was conducted to investigate effects of increasing doses: 0 (control), 6 (low), 12 (medium) and 24 (high) mg/g DM of ZADO^® enzyme preparation mixture (ENZ) on in vitro gas production (GP) and some ruminal fermentation parameters of the fibrous feeds Saccharum officinarum (leaves), Andropogon gayanus (leaves), Pennisetum purpureum (leaves) and Sorghum vulgare (straw). Rumen liquor was obtained from two Brown Swiss cows fitted with permanent rumen cannulae fed a total mixed ration of a 500:500 commercial concentrate and alfalfa hay ad libitum. The GP was recorded at 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h of incubation. After 96 h, the incubation was stopped and the pH of the mixture was determined and filtrate used to determine dry matter degradability (DMD), partitioning factor (PF₉₆), gas yield (GY₂₄), in vitro organic matter digestibility (OMD), metabolizable energy (ME), short chain fatty acids (SCFA), and microbial crude protien production (MCP). In general, the crude protein (CP) content of the fibrous feeds was low and ranged from 23 g/kg DM (S. officinarum) to 44 (A. gayanus). The fibre contents (i.e., NDFom and ADFom) were highest (P<0.05) in S. officinarum. Increasing ENZ dose linearly increased (P<0.05) GP of all fibrous feeds and had a quadratically increased (P<0.05) asymptotic gas production in P. purpureum and S. vulgare and rate of gas production in S. officinarum and S. vulgare. Addition of ENZ also quadratically increased (P<0.05) GP at all incubation times in S. officinarum and S. vulgare, and A. gayanus, but only at 72 h in A. gayanus. The parameters of ruminal fermentation of OMD, ME, GY₂₄ and SCFA linearly increased (P<0.05) and MCP linearly decreased (P<0.05) with the ENZ addition. Addition of enzyme affected ruminal fermentation of our feeds differently, mainly dependent on their fibre content, although dosage of enzyme was also important as impacts generally increased at higher dosages of ENZ.     

Effect of harvest date and nitrogen fertilization rate on the nutritive value of amaranth forage (Amaranthus hypochondriacus)

This study was conducted to assess effects of harvest date (i.e., 40 and 60 d after planting) and N fertilization rate (i.e., 120, 180, 240 kg N/ha) on the nutritive value of amaranth forage (Amaranthus hypochondriacus) using a factorial experiment with a randomized complete block design. The content of dry matter (DM), crude protein (CP), true protein (TP), ether extract (EE), water soluble carbohydrates (WSC), ash-free neutral detergent fiber (NDFom), ash-free acid detergent fiber (ADFom), lignin(sa), ash, Ca, P, Na, K, oxalic acid and nitrate were determined. Soluble CP (SP) and protein fractions non-protein N (A), true protein rapidly degraded in the rumen (B₁), true protein degraded in the rumen at a moderate rate (B₂), true protein associated with the cell wall and slowly degraded in the rumen (B₃) and acid detergent insoluble CP (C) were measured according to the Cornell Net Carbohydrate and Protein System. In vitro gas production (IVGP), OM disappearance (OMD) and NDFom disappearance (NDFD) were determined using a gas production technique. Results showed that the later harvest date increased (P<0.05) DM, EE, WSC, NDFom, ADFom, lignin(sa), B₃ and C; while CP, TP, ash, Ca, P, K, SP, A, B₁, B₂, nitrate, total and soluble oxalic acid, IVGP, b (i.e., gas production from the insoluble fermentable fractions at 120 h), c (i.e., rate of gas production during incubation), OMD and NDFD decreased (P<0.05). With increasing N fertilization rate, CP, TP, EE, P, nitrate, oxalic acid, SP, A, b, OMD and NDFD increased (P<0.05), however B₂ declined (P<0.05). Increasing N fertilization increased yield, CP concentration and nutrient digestibility. At 40 d after planting use of amaranth forage as a ruminant feed is limited due to its high nitrate content. However, at 60 d, although a depression in digestibility and CP content occurred, this forage has the potential as a ruminant feed due to the much lower nitrate levels.     

The application of the n-alkane technique for estimating the composition of diets consumed by equines and cattle feeding on upland vegetation communities

The application of n-alkanes as faecal markers to estimate diet composition of equines and cattle was studied. Twelve mature crossbreed mares (385 ± 47 kg live weight – LW) and six adult non-lactating cows (499 ± 36 kg LW) of Asturiana de los Valles breed were divided in groups of three animals (H1, H2, H3 and H4 in equines and C1 and C2 in cattle) and housed in individual stalls. Animals received a daily total amount of 1.0 kg DM/100 kg LW of different experimental diets composed of herbaceous (Lolium perenne L.) and woody species (Ulex gallii Planchon and heather: Erica spp. and Calluna vulgaris L.). Diet composition varied among treatments: H1 and C1 – L. perenne as sole component; H2 and C2 – L. perenne (0.70) and heather (0.30); H3 – L. perenne (0.70) and U. gallii (0.30); H4 – L. perenne (0.40), heather (0.30) and U. gallii (0.30).     

Nutrient composition and in vitro ruminal fermentation of tropical legume mixtures with contrasting tannin contents

Various combinations of a low-tannin herbaceous legume (Vigna unguiculata) and foliage of tanniniferous shrub legumes (Calliandra calothyrsus, Flemingia macrophylla and Leucaena leucocephala) or a low-tannin shrub legume (Cratylia argentea), all mixed together with a low-quality tropical grass (Brachiaria humidicola), were tested in vitro for differences in the effects on ruminal fermentation. Two experiments with the gas transducer technique were carried out, where each forage mixture was tested either with or without polyethylene glycol in order to be able to identify tannin-related effects (n = 3). In Experiment 1, a stepwise replacement of V. unguiculata by C. calothyrsus (5:0, 4:1, 3:2, 2:3, 1:4, 0:5) at a legume proportion of 1/3 or 2/3 in the mixture was evaluated. Together with two grass-alone and four pure legume treatments this added up to 30 treatments. In Experiment 2, V. unguiculata was gradually replaced by each of the four shrub legumes (3:0, 2:1, 1:2, 0:3) in grass–legume ratios of 2:1, adding up, together with two grass-alone treatments, to 28 treatments. When added alone, V. unguiculata resulted in high fermentative activity as measured by gas production and kinetics as well as low proportion of undegraded crude protein. When V. unguiculata was replaced by the low-tannin C. argentea in Experiment 2, there was no noticeable difference (P>0.05) in fermentative activity. In both experiments, the effect of the substitution of V. unguiculata by tanniniferous shrub legumes resulted in a declining gas production and an increasing proportion of undegraded crude protein (P<0.001). However, the extent of these changes depended on the level of replacement and the shrub legume species (P<0.001). The results of Experiment 2 illustrate that this was the consequence not only of different tannin contents (less adverse effects with L. leucocephala than with C. calothyrsus) but also differences in the chemical properties of the tannins present in these shrub legume species (much less adverse effects with L. leucocephala than with F. macrophylla despite similar tannin contents). Furthermore these results indicate that, once the extent of the effects of a tanniniferous legume is known, one may calculate the maximal level of replacement of a low-tannin legume in a grass diet possible without negative effects on ruminal fermentation. This allows to improve dry season grass-based diets with as few as possible of the expensive and less well growing low-tannin legume.     

Chemical composition, intake by sheep, and in situ disappearance in cannulated cows of bermudagrass hayed at two moisture concentrations and treated with a non-viable Lactobacillus-lactic acid preservative

Bermudagrass [Cynodon dactylon (L.) Pers.] is commonly used for grazing and haying in the southern USA, but hay curing can be challenging due to frequent rainfall events during spring and early summer. An existing stand of ‘Greenfield’ bermudagrass was divided into 12 plots using a randomized complete block design with a 2×2 factorial treatment arrangement to evaluate the influence of a non-viable Lactobacillus-lactic acid preservative and moisture concentration at baling on chemical composition, intake by sheep, and in situ disappearance in cattle. At time of mowing, half of the plots in each block were either spray-treated (T) or not treated (U) with 81 mL/t forage dry matter (DM) of the preservative solution. Hay was then baled at target moisture concentrations of either 174 g/kg DM (L) or 267 g/kg DM (H). Maximum temperature and heating degree days were greater (P<0.05) from H compared with L during the 42-d storage period. An interaction between spray and moisture treatments tended (P<0.10) to affect recovery of DM; recoveries for LT (0.992) differed (P<0.10) from HT (0.913), but LU and HU were intermediate between the spray-treated hays, and did not differ from either (P>0.10). Post-storage nutritive value was largely influenced by moisture treatments only. Intake and digestibility, and in situ DM disappearance of these same hays were determined using 16 wether lambs (43 ± 3.7 kg initial BW), or six ruminally cannulated cows (617 ± 3.5 kg initial BW), respectively. Dry matter intake by sheep was not affected by either treatment factor (P>0.05), but DM digestibility and digestible DM intake were greater (P<0.05) from U compared with T. The in situ immediately soluble DM portion was greater from (P<0.05) L compared with H, but the reverse was true for the potentially degradable DM fraction. The lag time tended (P<0.10) to be greater from H compared with L. Treating bermudagrass with a non-viable Lactobacillus acidophilus-lactic acid spray product at time of baling may not offset the negative effects on forage quality and digestibility of baling bermudagrass hay at excessive moisture concentrations.     

Effect of feed restriction or feeding high-fibre diet during the rearing period on body composition, serum parameters and productive performance of rabbit does

This study evaluated the effect of different feeding regimes from 11 weeks of age to first parturition on feed intake, leptin, non-esterified fatty acids (NEFA) and total protein serum levels, as well as productive performance in young rabbit does. In addition, body composition was estimated by bioimpedance analysis. Thirty-six 11-week-old does were randomly distributed in three groups. The AL-C group was fed ad libitum a control diet containing 350 g neutral detergent fibre (aNDFom)/kg, 11.6 MJ digestible energy (DE)/kg and 173 g crude protein (CP)/kg, and the does were inseminated at 16 weeks of age. The R-C group was fed 150 g/d of the same diet until 16 weeks of age, one week before artificial insemination (AI) at 17 weeks of age, and then fed ad libitum. The AL-F group was fed a diet containing 475 g aNDFom/kg, 9.4 MJ DE/kg and 174 g CP/kg ad libitum, and was inseminated at 17 weeks of age. During rearing (11-16 weeks), does in the R-C group had the lowest DE (1.54 MJ/d; P<0.003) and digestible protein (DP, 17.9 g/d; P<0.001) intake, as well as the lowest protein (172 g/kg; P<0.05) and energy (5.9 MJ/kg) body contents, leptin concentration at 16 weeks of age (2.48 ng/ml; P<0.001) and fertility (P<0.02) at first AI. Daily feed intake during pregnancy and lactation, as well as prolificacy and litter weight, were similar among groups. The highest percentage of body fat was observed for all the does when were inseminated for the first time (135 g/kg; P<0.001), consistent with the highest leptin (4.48 ng/ml; P<0.001) and total protein serum levels (6.87 g/dl; P<0.001) at this time. Serum NEFA level around parturition was higher (P<0.05) in groups AL-C and R-C (1.11 and 0.85 mmol/l) than in group AL-F (0.71 mmol/l), suggesting a lower lipid mobilization that tended to improve fertility rate for AL-F does on day 11 post-parturition (P<0.09). In conclusion, feed restriction during the rearing period delays reproductive development in young rabbits. In nulliparous does, ad libitum feeding during rearing with a high-fibre diet allows a similar productive performance to that of feeding with a less fibrous diet. Nevertheless, the use of high fibre diets during rearing does not affect feed intake throughout the first pregnancy and lactation.     

Nutritional characterization of Mucuna pruriens: 1. Effect of maturity on the nutritional quality of botanical fractions and the whole plant

This study was conducted to determine the stage of maturity at which the dry matter (DM) yield and nutritive value of velvet bean (Mucuna pruriens) is optimized. Mucuna was harvested at 77, 110 and 123 days after planting (DAP) from quadruplicate 5 m × 1 m plots within each of 6 blocks. At each DAP, DM yield, chemical composition, botanical composition, in vitro rumen fluid-pepsin DM digestibility (IVDMD) and concentrations of total polyphenols, l-dopa and tannins were determined on the whole plant and botanical fractions. Whole-plant Mucuna DM yield increased (P<0.01) linearly with maturity; proportions of leaves and stems decreased linearly (P<0.01), whereas proportion of pods increased (P<0.01). Concentrations of neutral-detergent fiber (aNDF) in whole plant, leaf, and stem increased (P<0.05), or tended (P<0.10) to increase linearly with maturity, as did the acid-detergent fiber concentration of leaves and stems. Maturity decreased (P<0.05) ether extract concentrations of leaves linearly, and stems quadratically, but increased (P<0.05) whole-plant and pod starch concentrations. Pods contained relatively high concentrations of lysine, histidine, phenylalanine, aspartate, glutamate, leucine, isoleucine, and valine, but low concentrations of methionine and cystine. The essential amino acid index did not vary with maturity. Most minerals in Mucuna are concentrated in the leaves and the whole plant contains sufficient Ca, P, K, Mg, Fe, Cu, Na, Mo, Mn, and Zn for growing sheep, although their bioavailability of these minerals is unknown. Total polyphenol concentration quadratically (P<0.01) increased with maturity in the whole plant, tended to increase (P<0.10) in pods, linearly (P<0.01) decreased in stems and fluctuated in leaves. Maturity quadratically increased l-dopa concentration of the whole plant (P<0.05) and stems (P<0.01), but did not affect those of leaves and pods. Maturity quadratically increased (P<0.05) total tannin concentration in the whole plant, but decreased (P<0.10) that of pods. The l-dopa was concentrated in the seeds and pods of mature (110–123 DAP) plants, but tannins were concentrated in leaves and stems. Whole-plant IVDMD was not affected by maturity, but digestible DM yield linearly (P<0.01) increased with increasing DM yield. There was a 2-week harvest window (110–123 DAP) during which whole-plant crude protein and IVDMD remained unchanged. Nevertheless, harvesting at 123 DAP gave the best combination of biomass yield and nutritive value.     

Safflower seeds in corn silage and alfalfa hay based early lactation diets: A practice within an optimum forage choice

Safflower seed (SS), Carthamus tinctorius L., has the highest concentration of linoleic acid among 80 oilseeds. It was hypothesized that an Iranian variety of SS can be effectively fed with cottonseeds (CS) to maintain feed intake, energy metabolism and productivity of early lactation cows under negative energy balance. Our objective was to determine effects of feeding diets containing 100 g whole CS with (1) no SS (SS0), (2) 75 g CS + 25 g SS (SS25), or (3) 50 g CS + 50 g SS (SS50), per kg of dietary DM, on feed intake, rumen fermentation, blood metabolites and milk production of early lactation cows fed diets based on a uniform mixtures of alfalfa hay and corn silage. Nine multiparous early lactation Holstein cows (46 ± 7 d in milk) were used in a replicated 3 × 3 Latin square design study with three 21-d periods. Each period had 14 d of adaptation and 7 d of data collection. Dietary inclusion of SS did not affect (P>0.10) DM intake, rumen pH and concentrations of ammonia and VFA, blood concentrations of insulin, non-esterified fatty acids, urea and triglycerides, and milk production. Adding SS linearly reduced blood glucose (P=0.05) and beta-hydroxybutyric acid (P<0.05), and increased blood total cholesterol (P<0.01) and low-density lipoproteins (P<0.05) concentrations. Results demonstrated that SS as an economical and rich source of essential fatty acids can be included up to 50 g/kg of dietary DM alongside CS for early lactation cows without affecting feed intake while maintaining rumen fermentation, peripheral energy supply and milk production.     

Chemical composition and rumen degradability of three corn hybrids treated with insecticides against the European corn borer (Ostrinia nubilalis)

This experiment determined the chemical composition, rumen degradability (aNDF in stalks and starch in kernels) and in vitro gas production of kernels from three corn hybrids treated (TT) or not treated (control, CTR) with insecticides against the European corn borer (ECB, Ostrinia nubilalis). Two whole-plant silage hybrids belonging to the FAO rating 600 and 700 maturity class (S600 and S700, respectively) and one selected for grain production (G600, FAO rating 600, Dekalb-Monsanto Agricoltura S.p.A., Lodi, Italy) were sown in two main plots (TT and CTR) of an experimental field. Two subsequent treatments of pyrethroids (25 and 1.2 g/ha of cyfluthrin and deltamethrin, respectively) were applied to the TT plots. The insecticide treatment reduced the number of damaged plants (4.5 broken plants/plot versus 0.3 broken plants/plot, P<0.01) and increased the total grain yield by 11% (13.8 t/ha versus 12.4 t/ha), while hybrids did not differ. ECB larvae which bored into the stalk tunnels modified the chemical composition of stalks and kernels. In stalks, total sugars content (i.e. glucose, fructose, sucrose) was about twice that in TT versus CTR plants (123 g/kg versus 60 g/kg DM, P<0.01), while aNDF content was higher in CTR stalks (765 versus 702 g/kg DM, P<0.01). DM degradability after 48 h of incubation of stalks was higher in TT than in CTR, both in vitro (0.360 versus 0.298, P<0.01) and in situ (0.370 versus 0.298, P<0.05), while there were no differences in aNDF degradability. Kernels from TT plots contained less DM (615 g/kg versus 651 g/kg, P<0.01) and more CP (84 g/kg and 78 g/kg DM, P<0.05) than those from CTR plots, while in situ rumen starch disappearance and in vitro gas production were similar. Corn hybrid selected for yield of grain (G600) differed from S600 and S700 due to a higher (P<0.01) content of aNDF, ADF and lignin(sa) in the stalks, and a higher starch content (696 g/kg versus 674 and 671 g/kg DM, P<0.01) and CP (87 g/kg versus 77 and 76 g/kg DM, P<0.05) in grain. The G600 hybrid produced stalks with a lower (P<0.01) aNDF rumen degradability than the S600 and S700.     

Assessment of changes in DNA methylation by methylation-sensitive amplification polymorphism in Jatropha curcas L. subjected to salinity stress

The present study assesses the changes in DNA methylation in leaf and root tissues of Jatropha curcas L., induced by salinity stress using methylation sensitive amplification polymorphism (MSAP) markers. Seedlings of 21 days (d) grown under controlled conditions were subjected to 0–100 mM salinity treatment for 24 h (1 d). Immediate changes in DNA methylation and polymorphism in methylated DNA in whole genome of both leaves and roots were assessed using 10 selective combinations of MSAP primers. In root and leaves 70.06% and 57.89% methylation was observed respectively. Similarly 67.22% and 71.21% polymorphism was observed in methylated DNA from root and leaf tissues respectively. Compared with control, the percentage of methylation and methylation polymorphism in roots of plants under different dosages of salinity was found in the order of 50 mM < 25 mM = 100 mM < 75 mM and 75 mM < 25 mM < 50 mM < 100 mM respectively. Similarly percentage of methylation and methylation polymorphism in leaves of plants treated with different levels of salinity was found in order of 75 mM < 25 mM < 50 mM < 100 mM and 50 mM < 25 mM < 100 mM < 75 mM respectively. The MSAP analysis showed that under salt stress homologous nucleotide sequences in genome from control and salt treated plants of J. curcas showed different patterns of methylation; which suggest that these fragments probably play an important role to induce immediate adaptive responses in Jatropha under salinity stress.     

Effects of hybrid and maturity on performance and nutritive characteristics of forage maize at high latitudes, estimated using the gas production technique

We evaluated effects of hybrid and advancing plant maturity on performance, chemical composition, and nutritional characteristics of whole plant forage maize as well as the relative contributions of its plant fractions at high latitudes. Three maize hybrids, Avenir (FAO 180), Isberi (FAO 190) and Burli (FAO 210), were grown in southern Sweden in a field experiment with a replicated complete randomized block design. Plants were harvested four times during maturation, and dry matter (DM) yield and DM as a proportion of fresh weight were recorded. Whole plants were separated into four morphological fractions representing stems, leaves, kernels and cobs and the contribution of each fraction to the DM of the whole plant was estimated. Plant material was subjected to chemical analysis followed by measurement of in vitro gas production (GP) in buffered rumen fluid and finally, by calculation of in vitro true digestibility of organic matter (OM) and neutral detergent fibre (aNDFom). The GP profiles were fitted to a first order kinetic model with a discrete lag. Parameters describing the GP profiles were used in a recently developed mechanistic two compartment rumen degradation model to estimate in vivo OM digestibility (OMD) and first order rate of degradation in the rumen. Hybrids were compared and effects of maturity were assessed by analysis of variance using DM concentration as covariate. There were differences (P<0.05) among the hybrids in DM yield and relative contributions of the plant fractions. Differences (P<0.05) in modelled in vivo digestibility of OM and rates of degradation also occurred among hybrids. Increased maturity caused a reduction in in vitro digestibility of aNDFom in all plant fractions (P<0.05), but increased the rate of rumen degradation of OM in the whole plant as evaluated from GP results. The DM yield had a quadratic relationship with increasing maturity, with maximum yield at a DM concentration of about 370 g/kg. In vitro GP can describe the nutritive characteristics of forage maize in relation to advancing maturity and increased maturity affected agronomic performance and plant composition as well as the nutritive characteristics of hybrids.     

Genetic effects of root extracts of Glycyrrhiza glabra L. in different test systems

The antimutagenic and geroprotective activities of licorice (Glycyrrhiza glabra) root extracts were established in cells of plant test systems of Allium fistulosum L., Allium cepa L., and Vicia faba L. and of laboratory animals (Vistar rats). The prospects for the practical application of licorice Glycyrrhiza glabra root extracts as antimutagenic substances are discussed.     

Seasonal variations in the chemical composition and dry matter degradability of exclosure forages in the semi-arid region of northern Ethiopia

The present study was conduced at two sites (Tembien and Wukro) in the semi-arid region of Tigray in northern Ethiopia to investigate the seasonal dynamics in the chemical composition and dry matter digestibility of grass and browse species of exclosures. The browse species studied in Tembien and Wukro had a mean crude protein (CP) value of 166 and 117 g/kg dry matter (DM), respectively. The mean in vitro dry matter digestibility (IVDMD) coefficient and predicted metabolizable energy (ME) density of the browse species were 0.72 and 9.83 MJ/kg DM, respectively at Tembien, 0.62 and 8.38 MJ/kg DM, respectively, at Wukro. Neutral detergent fibre (NDF) and acid detergent fibre (ADF) values of the browse species varied from 192 to 437 and 127 to 391 g/kg DM, respectively. Acid detergent lignin (ADL) values ranged from 36 to 190 g/kg DM. The mean CP of the grass species in Tembien and Wukro during the long rainy season was 76 and 73 g/kg DM, respectively and values declined below a critical maintenance level during the dry and short rain seasons. Mean IVDMD and ME values for the two sites were 0.41 and 0.47, and 5.38 and 6.11 MJ/kg DM, respectively. The NDF, ADF, and lignin values of the grass species were generally above 700, 400, and 70 g/kg DM, respectively. The CP, IVDMD and ME values of the mixed grass samples differed (P<0.05) among harvesting months and values ranged from 20 to 103 g/kg DM, 0.47 to 0.72 MJ/kg DM, and 6.16 to 9.91 MJ/kg DM, respectively. The browse species could be used as useful dry season protein supplements to the N deficient native grass species. Especial emphasis should be given to propagate Maerua angolensis and Cadaba farinosa at community nursery sites. Harvesting in September, rather than the current extended harvest period that took place in October and November, can considerably improve the feeding value of native grass hay for smallholder ruminant production systems.     

Optimisation of a microwave pretreatment of wheat straw for methane production

This study aims at the optimisation of a microwave pretreatment for wheat straw solubilisation and anaerobic biodegradability. The maximum yield of methane production was obtained at 150 °C with an improvement of 28% compared to an untreated sample. In addition, at this temperature, the time to reach 80% of the methane volume obtained from untreated straw was about 35%. The study of ramp time and holding time at targeted temperature showed that they had no improvement effect. Thus, the best conditions are the highest heating rate for a final temperature 150 °C without any holding time. The reading of energy consumed by pretreatment and energy overproduced by pretreated samples showed that increasing tVS amount and heating rate led to a saving of energy consumption. Nevertheless, to obtain a positive energy balance, a microwave device should consume less than 2.65 kJ/g_tVS.     

Temperate forages ensiled with molasses or fresh cheese whey: Effects on conservation quality, effluent losses and ruminal degradation

Effects of ensiling and adding molasses or increasing levels of fresh cheese whey on the conservation and rumen degradability of temperate pasture silages were evaluated. Forage from three paddocks of mixed grass and legume pastures was used to make 45 silages (15 silages per paddock) with 5 treatments, including silage without additives (control), silage with 15 g/kg dehydrated molasses and silage with 20, 50 and 100 g/kg fresh cheese whey. The chemical composition and fermentation quality (i.e., pH, ammonia N, loss of dry matter (DM) and neutral detergent fibre (NDF), effluent production) of the silages were determined. Fresh and ensiled materials were evaluated for in situ rumen degradability. Ensiling reduced DM and NDF rumen degradability (P<0.01). When additives were employed, the reduction of DM degradability of the silages decreased (P≤0.03). Addition of molasses led to the lowest pH (P<0.01) and DM losses (P<0.01), and highest DM degradability (P<0.01). The conservation and DM degradation results of dried molasses silage was superior to those of fresh whey silages. In general, an increase in the level of whey increased DM degradability (P≤0.03), but linearly increased effluent production (P<0.01) and losses (P<0.01).     

Optimization of cellulase production by a brown rot fungus Fomitopsis sp. RCK2010 under solid state fermentation

Culture conditions for enhanced cellulase production from a newly isolated brown rot fungus, Fomitopsis sp. RCK2010 were optimized under solid state fermentation. An initial pH of 5.5 and moisture ratio of 1:3.5 (solid:liquid) were found to be optimal for maximum enzyme production. Of the different carbon sources tested wheat bran gave the maximum production of CMCase (71.526 IU/g), FPase (3.268 IU/g), and β-glucosidase (50.696 IU/g). Among the nitrogen sources, urea caused maximum production of CMCase (81.832 IU/g), where as casein and soyabean meal gave the highest FPase (4.682 IU/g) and β-glucosidase (69.083 IU/g) production, respectively. Among amino acids tested glutamic acid gave the highest production for CMCase (84.127 IU/g); however 4-hydroxy-l-proline stimulated maximum FPase production (6.762 IU/g). Saccharification of pretreated rice straw and wheat straw by crude enzyme extract from Fomitopsis sp. RCK2010 resulted in release of 157.160 and 214.044 mg/g of reducing sugar, respectively.