Carbohydrates and protein digestive traits in beef cattle grazing fertilised or mixed tropical pasture.

This study was performed to investigate the nitrogen (N) and carbohydrate digestive traits of grazing heifers. The experiment was carried out at the Federal University of Lavras. The treatments were a Marandu palisadegrass (Urochloa brizantha [Syn, Brachiaria brizantha] Stapf. A. Rich. cv. Marandu) monoculture fertilised with 150 kg N/[ha ∙ year] (FP) or Marandu palisadegrass mixed pasture with forage peanut (MP). The pastures were grazed by six rumen-cannulated zebu heifers. A double cross-over design was used in four periods. Nutritive value, intake and apparent digestibility of forage, ruminal traits and kinetics and N balance were evaluated. Apparent total-tract digestibility of dry matter (DM) and neutral detergent fibre (NDF) were greater for FP than for MP. There was no effect in apparent total-tract digestibility of N. The estimated intestinal digestibility of nutrients was greater on MP than FP. Even though N intake and faecal N output were greater on MP than FP, there was no effect in urine N output. The N balance tended to be greater on MP than FP. The forage peanut, which contains condensed tannins, decreased ruminal fibre degradation, apparent digestibility and ruminal protein degradation, increased N flow from the rumen. Inclusion of forage peanut in the mixed pasture decreased the ruminal fibre degradability but increased N retention by the animals.

Greenhouse Gas Emissions and Crossbred Cow Milk Production in a Silvopastoral System in Tropical Mexico.

In Mexico, pasture degradation is associated with extensive pastures; additionally, under these conditions, livestock activities contribute considerably to greenhouse gas (GHG) emissions. Among the options to improve grazing systems and reduce GHG emissions, silvopastoral systems (SPS) have been recommended. The objectives of this work were to quantify the N outflow in a soil-plant-animal interface, as well as the CH4 emissions and milk production in an SPS with woody legumes (Leucaena leucocephala) that is associated with stargrass (Cynodon nlemfuensis). This was then compared with stargrass in a monoculture system (MS) in the seasons (dry and rainy period) over a two-year period. Dung was collected from the animals of each of the grazing systems and applied fresh to the land plots. Fresh dung and urine were collected from the cows of each grazing system and were applied to the experimental plots. In addition, the soil CH4 and N2O contents were measured to quantify the emissions. Average milk yield by seasons was similar: MS (7.1 kg per animal unit (AU)/day-1) and SPS (6.31 kg per AU/day-1). Cows in the MS had a mean N intake of 171.9 g/UA day-1 without seasonal variation, while the SPS animals' mean N intake was 215.7 g/UA day-1 for both seasons. For the urine applied to soil, the N2O outflow was higher in the MS (peak value = 1623.9 µg N-N2O m-2 h-1). The peak value for the SPS was 755.9 µg of N-N2O m-2 h-1. The N2O emissions were higher in the rainy season (which promotes denitrification). The values for the feces treatment were 0.05% (MS) and 0.01% (SPS). The urine treatment values were 0.52% (MS) and 0.17% (SPS). The emissions of CH4 showed that the feces of the SPS systems resulted in a higher accumulation of gas in the rainy season (29.8 g C ha-1), followed by the feces of the MS system in the dry season (26.0 g C ha-1). Legumes in the SPS helped to maintain milk production, and the N2O emissions were lower than those produced by the MS (where the pastures were fertilized with N).

Bradyrhizobium occurrence in nodules of perennial horsegram.

The introduction of a forage legume into a tropical pasture should decrease the need for N fertilizer, provided biological N2 fixation (BNF) contributes enough to compensate for exported N. Macrotyloma axillare (perennial horsegram) is a suitable legume for composing mixed pastures, and our hypothesis is that the isolation of indigenous rhizobia from roots and rhizosphere is the way of achieving an efficient inoculant to maximize BNF to the legume. Nodules and rhizosphere soil taken from M. axillare grown in a mixed pasture with palisade grass were sampled and used in a trap host assay using Leonard jars containing a mixture of vermiculite and sand. A total of ten bacteria were initially isolated using this technique. The isolates were then used in two experiments to evaluate the inoculation responses on the perennial horsegram in greenhouse conditions to which nodulation, plant growth, and shoot N accumulation were measured. Phylogenetic analyses based on 16S rRNA and recA placed all strains within genus Bradyrhizobium, some of them not previously described. The best strain provided more than 120 nodules and more than 65 mg of nodules per plant. Strain BR14182 was considered the most promising given the high dry matter and N accumulation in plant shoots. This study provides the first analysis of Bradyrhizobium diversity nodulating M. axillare in Brazil and provided evidence of the role of inoculation in incrementing the plant-rhizobium symbiosis in a forage legume.

Could forage peanut in low proportion replace N fertilizer in livestock systems?

Palisadegrass [Urochloa brizantha (Hochst. ex A. Rich.) R. D. Webster cv. Marandu] is widely used in Brazil and is typically managed with little or no N fertilizer, which often leads to pasture decline in the long-term. The current relationship between beef price and fertilizer cost in Brazil does not favor fertilizer use in pastures. Legume inclusion is an alternative to adding fertilizer N, but often legumes do not reach a significant proportion (> 30%) in pasture botanical composition. This study evaluated herbage responses to N inputs and pasture species composition, under intermittent stocking. Treatments included palisadegrass-forage peanut (Arachis pintoi Krapov. & W.C. Greg. cv. Amarillo) mixture (mixed), unfertilized palisadegrass (control), and palisadegrass fertilized with 150 kg N ha-1 yr-1 (fertilized). Treatments were applied over two rainy seasons with five growth cycle (GC) evaluations each season. Response variables included herbage biomass, herbage accumulation, morphological components, total aboveground N of forage peanut (TAGNFP), and contribution of biological N2 fixation (BNF). Herbage biomass was greater for fertilized palisadegrass [5850 kg dry matter (DM) ha-1] than for the palisadegrass-forage peanut mixture (3940 kg DM ha-1), while the unfertilized palisadegrass (4400 kg DM ha-1) did not differ from the mixed pasture. Nitrogen fertilizer increased leaf mass of palisadegrass (2490 kg DM ha-1) compared with the control and mixed treatments (1700 and 1310 kg DM ha-1, respectively). The contribution of BNF to the forage peanut ranged from 79 to 85% and 0.5 to 5.5 kg N ha-1 cycle-1. Overall, benefits from forage peanut were minimal because legume percentage was less than 10%, while N input in the system by N-fertilizer increased palisadegrass herbage biomass.

Nitrogen supply and rainfall affect ammonia emissions from dairy cattle excreta and urea applied on warm-climate pastures.

Cattle excreta and nitrogen (N) fertilizer deposited on tropical grasslands are important sources of ammonia (NH3 ) emission. We conducted three field trials (wet, intermediate, and dry conditions) to quantify NH3 emissions from urea fertilizer and simulated excretions of heifer urine and dung on warm-climate grasslands in Brazil. Heifer excreta were derived from pastures of palisadegrass [Urochloa brizantha (Hochst. ex A. Rich.) R. D. Webster 'Marandu'] under three forms of N supply (without or with N fertilization [0 or 150 kg N ha-1  yr-1 ] or mixed with forage peanut [Arachis pintoi 'Amarillo']). Cumulative NH3 -N emissions across rainfall conditions were 7.6-16.6% (mean, 11.7%) for urine, 1.4-2.9% (mean, 2.0%) for dung, and 11.2-20.5% (mean, 14.8%) for urea. Ammonia loss from urine was significantly greater than from dung under all rainfall conditions. Emission from urine and dung differed from those when urea was applied on palisadegrass. There were greater NH3 emissions from urine in the wetter times of the year. Heifer excreta from N-fertilized pasture had greater NH3 emission than excreta from the grass-legume mixture and unfertilized palisadegrass. Urea applied on palisadegrass presented greater NH3 emissions in wet rainfall conditions compared with dry conditions but did not differ from intermediate conditions. Our study showed that N-fertilized systems increase N losses as NH3 emission from excreta, and emissions from urea fertilizer must be included in this system. Heifer excreta and urea fertilizer deposited on warm-climate grasslands increased the NH3 emissions mainly under wet conditions.

Litter decomposition and nutrient release dynamics of leaves and roots of the babassu palm in eastern Amazonia

The ruderal babassu palm (Attalea speciosa) is expanding on large areas of degraded Amazon landscapes. Decomposition of leaves and roots is in the center of plant:soil interactions. We evaluated decomposition and nutrient concentrations of leaves and fine roots of babassu in comparison with two exotic reference species, Acacia mangium (slow degradability) and Leucaena leucocephala (fast degradability), in a 138-day litterbag assay carried out in secondary forest stands of different age and babassu abundance. We chose 4-mm over 2-mm mesh litterbags based on a pilot study. Babassu leaves degraded slower than leaves of A. mangium and L. leucocephala, and also had lower nitrogen, phosphorus and calcium concentrations in all stages of decomposition. By contrast, potassium concentrations in babassu leaves were higher than in both reference species at 0 and 50 days. Roots of all three species decomposed slower than leaves. Compared to the leaves, both biomass loss and nutrient concentrations differed less between babassu and reference-species roots, except for lower nitrogen concentration in babassu roots. Leaf-litter decomposition of all three species was significantly faster in old than in young secondary forest, suggesting an acceleration of decomposition along succession. Babassu leaves decomposed faster in old babassu-dominated than non-dominated secondary forest, pointing to the existence of specialized decomposer communities in babassu-dominated stands. (AU)

Increase in Milk Yield from Cows through Improvement of Forage Production Using the N2-Fixing Legume Leucaena leucocephala in a Silvopastoral System.

The objective was to evaluate milk production, N2-fixation and N transfer, forage yield and composition (under two cutting intervals) in a silvopastoral system (SPS) with Leucaena leucocephala-Megathyrsus maximus and M. maximus-monoculture (MMM) with crossbred cows in a completely randomized design. Forage yield in the SPS was 6490 and 6907 kg DM ha-1 for cutting intervals (CI) of 35 and 50 days. Forage yield for the MMM was 7284 and 10,843 kg DM ha-1, and forage crude protein (CP) was 29.0% and 26.1% for L. leucocephala, harvested at 35 and 50 days, respectively. CP for the associated M. maximus was 9.9% and 7.8% for CI 35 and 50 days, respectively, and for MMM was 7.4% and 8.4%, harvested at 35 and 50 days. Milk production was 4.7 kg cow-1 day-1 for cows grazing MMM and 7.4 kg cow-1 day-1 under SPS. Nitrogen fixation in L. leucocephala (%Ndfa) was estimated to be 89% and 95%, at 35 and 50 days, with an N2 transfer to the associated grass of 34.3% and 52.9%. SPS has the potential to fix and transfer important amounts of N2 to the associated grass, and increase forage CP content and milk production.

Methane production and nitrogen balance of dairy heifers grazing palisade grass cv. Marandu alone or with forage peanut.

Livestock production systems are an essential agribusiness activity in Brazil, but a critical challenge of Brazilian farmers is to maintain the equilibrium of the ecosystem, using herbage resources efficiently with a minimum impact on the environment. Nitrogen (N) fertilization and the inclusion of forage legumes into tropical grass pastures are management strategies which increase the productivity and nutritive value of pastures and may also affect methane (CH4) production by ruminants. The objective of this study was to examine the effects of either fertilizing palisade grass pastures with N or including the forage peanut (Arachis pintoi) into grass pastures on enteric CH4 emission, microbial protein production in the rumen via purine derivatives in the urine, and N balance. Twenty-one nonlactating crossbred dairy heifers were used in a completely randomized design with 3 treatments. The treatments consisted of pastures of palisade grass without N fertilization (control), fertilized with urea (fertilized), and palisade grass mixed with forage peanut (mixed). Seven animals (replications) were used to evaluate dry matter intake, digestibility, CH4 emission, urea, purine derivatives, and volume of urine, and N ingestion and excretion. Four paddocks (replications) were used to measure herbage mass; morphological, botanical, and chemical composition of herbage; and herbage allowance. The CH4 emissions were determined using the sulfur hexafluoride (SF6) tracer gas technique. The efficiency of N utilization (ENU) was calculated using the N balance data. Crude protein (CP) concentration of herbage increased with fertilization or legumes inclusion (P < 0.0001) while neutral detergent fiber (NDF) concentration decreased (P = 0.0355). The leaf allowance was higher in the fertilized treatment (P = 0.0294). Only uric acid excretion increased with N fertilization (P = 0.0204). The ENU was not affected by fertilized or mixed compared to control and averaged 55% (P = 0.8945). The enteric CH4 production was similar between treatments and averaged 129 g/d (P = 0.3989). We concluded that the changes in chemical composition of herbage provided by N fertilization or the inclusion of the legume showed no reduction in enteric CH4 emissions, but the ENU was more significant than previous studies with palisade grass, suggesting that different management strategies might alter the ENU under grazing conditions.

Bacteria related to Bradyrhizobium yuanmingense from Ghana are effective groundnut micro-symbionts.

The identification of locally-adapted rhizobia for effective inoculation of grain legumes in Africa's semiarid regions is strategic for developing and optimizing cheap nitrogen fixation technologies for smallholder farmers. This study was aimed at selecting and characterising effective native rhizobia, from Ghanaian soils for groundnut (Arachis hypogaea L.) inoculation. From surface-disinfected root nodules of cowpea and groundnut plants grown on farmers' fields, 150 bacterial isolates were obtained, 30 of which were eventually found to nodulate groundnut plants. After testing the symbiotic potential of these isolates on groundnut on sterilized substrate, seven of them, designated as KNUST 1001-1007, were evaluated in an open field pot experiment using 15N-labelled soil. Although 15N dilution analyses did not indicate differences among treatments in the proportion of nitrogen (N) derived from the atmosphere (%Ndfa), all seven strains increased total N derived from N2 fixation by inoculated groundnut plants as compared to the non-inoculated control. Inoculation with KNUST 1002 led to total N accumulation as high as that of the groundnut reference strain 32H1. Genetic characterisation of the isolates by sequence analysis of 16S rRNA gene, 16S - 23S rRNA intergenic transcribed spacer (ITS) region and nodC gene revealed that isolates KNUST 1003 and 1007 were related to Rhizobium tropici, a common bean symbiont. The other five isolates, including KNUST 1002 belonged to the Bradyrhizobium genus, being closely related to Bradyrhizobium yuanmingense. Therefore, this study revealed novel native Ghanaian rhizobia with potential for the development of groundnut inoculants.

Effects of grazing management in brachiaria grass-forage peanut pastures on canopy structure and forage intake1.

Maintenance of mixed grass-legume pastures for stand longevity and improved animal utilization is a challenge in warm-season climates. The goal of this study was to assess grazing management on stand persistence, forage intake, and N balance of beef heifers grazing mixed pastures of Brachiaria brizantha and Arachis pintoi. A 2-yr experiment was carried out in Brazil, where four grazing management were assessed: rest period interrupted at 90%, 95%, and 100% of light interception (LI) and a fixed rest period of 42 d (90LI, 95LI, 100LI, and 42D, respectively). The LI were taken at 50 points at ground level and at 5 points above the canopy for each paddock using a canopy analyzer. For all treatments, the postgrazing stubble height was 15 cm. Botanical composition and canopy structure characteristics such as canopy height, forage mass, and vertical distribution of the morphological composition were evaluated pre- and post-grazing. Forage chemical composition, intake, and microbial synthesis were also determined. A randomized complete block design was used, considering the season of the year as a repeated measure over time. Grazing management and season were considered fixed, while block and year were considered random effects. In the summer, legume mass accounted for 19% of the canopy at 100LI, which was less than other treatments (a mean of 30%). The 100LI treatment had a greater grass stem mass compared with other treatments. In terms of vertical distribution for 100LI, 38.6% of the stem mass was above the stubble height, greater than the 5.7% for other treatments. The canopy structure limited NDF intake (P = 0.007) at 100LI (1.02% of BW/d), whereas 42D, 90LI, and 95LI treatments had NDF intake close to 1.2% of BW/d. The intake of digestible OM (P = 0.007) and the ratio of CP/digestible OM (P < 0.001) were less at 100LI in relation to the other treatments. The production of microbial N (P < 0.001) and efficiency of microbial synthesis (P = 0.023) were greater at 95LI and 90LI, followed by 42D and less at 100LI. Overall, the range from 90% to 95% of LI is the recommendation to interrupt the rest period, since this strategy enhanced community stability, forage intake, and nutritional value of the diet. Under on-farm conditions, brachiaria grass and forage peanut pastures should be managed at a range height of 24 to 30 cm.

Nitrous Oxide and Methane Fluxes Following Ammonium Sulfate and Vinasse Application on Sugar Cane Soil.

This study aimed to quantify nitrous oxide (N2O) and methane (CH4) emission/sink response from sugar cane soil treated with fertilizer nitrogen (N) and vinasse applied separately or in sequence, the latter being investigated with regard to the time interval between applications for a possible effect on emissions. The study was carried out in a traditional area of unburned sugar cane in São Paulo state, Brazil. Two levels of N fertilization (0 and 100 kg N ha(-1)) with no added vinasse and combined with vinasse additions at different times (100 m(-3) ha(-1) at 3 and 15 days after N fertilization) were evaluated. Methane and N2O fluxes were monitored for 211 days. On average, the soil was a sink for CH4, which was not affected by the treatments. Emissions of N2O were induced by N fertilizer and vinasse applications. For ammonium sulfate, 0.6% of the added N was emitted as N2O, while for vinasse, this ranged from 1.0 to 2.2%. Changes in N2O fluxes were detected the day after application of vinasse on the N fertilized areas, but although the emission factor (EF) was 34% greater, the EF was not significantly different from fertilizer N alone. Nevertheless, we recommend to not apply vinasse after N fertilization to avoid boosting N2O emissions.

Nitrogen-fixing legume tree species for the reclamation of severely degraded lands in Brazil.

The main challenges faced in the reclamation of severely degraded lands are in the management of the systems and finding plant species that will grow under the harsh conditions common in degraded soils. This is especially important in extremely adverse situations found in some substrates from mining activities or soils that have lost their upper horizons. Under these conditions, recolonization of the area by native vegetation through natural succession processes may be extremely limited. Once the main physical and chemical factors restrictive to plant growth are corrected or attenuated, the introduction of leguminous trees able to form symbioses with nodulating N2-fixing bacteria and arbuscular mycorrhizal fungi constitutes an efficient strategy to accelerate soil reclamation and initiate natural succession. These symbioses give the legume species a superior capacity to grow quickly in poor substrates and to withstand the harsh conditions presented in degraded soils. In this article we describe several successful results in Brazil using N2-fixing legume tree species for reclamation of areas degraded by soil erosion, construction and mining activities, emphasizing the potential of the technique to recover soil organic matter levels and restore ecosystem biodiversity and other environmental functions.

Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil.

*An extensive survey of nodulation in the legume genus Mimosa was undertaken in two major biomes in Brazil, the Cerrado and the Caatinga, in both of which there are high degrees of endemicity of the genus. *Nodules were collected from 67 of the 70 Mimosa spp. found. Thirteen of the species were newly reported as nodulating. Nodules were examined by light and electron microscopy, and all except for M. gatesiae had a structure typical of effective Mimosa nodules. The endosymbiotic bacteria in nodules from all of the Mimosa spp. were identified as Burkholderia via immunolabelling with an antibody against Burkholderia phymatum STM815. *Twenty of the 23 Mimosa nodules tested were shown to contain nitrogenase by immunolabelling with an antibody to the nitrogenase Fe- (nifH) protein, and using the delta(15)N ((15)N natural abundance) technique, contributions by biological N(2) fixation of up to 60% of total plant N were calculated for Caatinga Mimosa spp. *It is concluded that nodulation in Mimosa is a generic character, and that the preferred symbionts of Brazilian species are Burkholderia. This is the first study to demonstrate N(2) fixation by beta-rhizobial symbioses in the field.

Quantification of natural populations of Gluconacetobacter diazotrophicus and Herbaspirillum spp. in sugar cane (Saccharum spp) using differente polyclonal antibodies

The species Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN) Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay) can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 10(5) cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques.

Quantification of natural populations of Gluconacetobacter diazotrophicus and Herbaspirillum spp. In sugar cane (Saccharum spp.) Using differente polyclonal antibodies.

The species Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN) Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay) can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 10(5) cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques.