Forage peanut legume as a strategy for improving beef production without increasing livestock greenhouse gas emissions.

The transformation of pastures from a degraded state to sustainable productivity is a major challenge in tropical livestock production. Stoloniferous forage legumes such as Arachis pintoi (forage peanut) are one of the most promising alternatives for intensifying pasture-based beef livestock operations with reduced greenhouse gas (GHG) emissions. This 2-year study assessed beef cattle performance, nutrient intake and digestibility, and balance of GHG emissions in three pasture types (PT): (1) mixed Palisade grass - Urochloa brizantha (Hochst. ex A. Rich.) R.D. Webster (syn. Brachiaria brizantha Stapf cv. Marandu) and forage peanut (A. pintoi Krapov. & W.C. Greg. cv. BRS Mandobi) pastures (Mixed), (2) monoculture Palisade grass pastures with 150 kg of N/ha per year (Fertilised), and (3) monoculture Palisade grass without N fertiliser (Control). Continuous stocking with a variable stocking rate was used in a randomised complete block design, with four replicates per treatment. The average daily gain and carcass gain were not influenced by the PT (P = 0.439 and P = 0.100, respectively) and were, on average, 0.433 kg/animal per day and 83.4 kg/animal, respectively. Fertilised and Mixed pastures increased by 102 and 31.5%, respectively, the liveweight gain per area (kg/ha/yr) compared to the Control pasture (P < 0.001). The heifers in the Mixed pasture had lower CH4 emissions (g/animal per day; P = 0.009), achieving a reduction of 12.6 and 10.1% when compared to the Fertilised and Control pastures, respectively. Annual (N2O) emissions (g/animal) and per kg carcass weight gain were 59.8 and 63.1% lower, respectively, in the Mixed pasture compared to the Fertilised pasture (P < 0.001). Mixed pasture mitigated approximately 23% of kg CO2eq/kg of carcass when substituting 150 kg of N/ha per year via fertiliser. Mixed pastures with forage peanut are a promising solution to recover degraded tropical pastures by providing increased animal production with lower GHG emissions.

Effects of supplementation with corn distillers' dried grains on animal performance, nitrogen balance, and enteric CH4 emissions of young Nellore bulls fed a high-tropical forage diet.

The inclusion of corn-dried distillers' grains (DDG) could be an alternative supplement to increase animal performance, nitrogen efficiency usage (NEU), and decrease enteric methane (CH4) emissions. Our goal was to determine whether DDG could replace a traditional supplement (cottonseed meal) without affecting animal performance, N balance, and CH4 emissions. The experiment was conducted during the forage growing season (December to April), with 15 d adaptation, and a 112 d experimental period. The experimental design was completely randomized with four treatments: a mineral supplement (MS), cottonseed meal supplement (CS), 50% replacement of CS by DDG (50DDG), and 100% replacement of CS by DDG (100DDG). Cottonseed meal and DDG were used as protein supplement. A total of 12 paddocks, 3 per treatment, were used to measure forage mass: morphological and chemical composition of forage, forage allowance, and animal performance. Six animals per treatment were used to evaluate DM intake, digestibility, CH4 emissions, microbial protein production (MCP), and NEU of each treatment. Eighty-one Young Nellore bulls (48 testers, 12 per treatments and 33 adjusters) with initial BW of 255 ±â€¯5 kg (10-12 months old) were supplemented with each supplement type at a level of 0.3% of BW. Pasture management was continuous stocking with a variable stocking rate (put-and-take). Enteric CH4 was measured using the gas tracer technique. The MCP was quantified using purine derivatives and the NEU mass balance. No differences were found in nutrient intake (P > 0.228). Individual animal performance and gain per area were higher in the treatments with concentrates compared with that of MS; however, there was no difference among treatments CS, 50DDG, and 100DDG. The ADG was 0.83 for MS and 1.08 kg/animal/d when supplemented (P < 0.05). Gain per hectare was 709 kg/ha for MS and 915 kg/ha when supplemented with concentrates (P < 0.05). There was no difference in CH4 production among treatments that average 180 g/animal/d; however, CH4 per kg of gain was reduced with CS. The CH4 conversion factor averaged 5.91%. There was no difference in the synthesis of MCP and NEU. Corn DDG can replace 100% of cottonseed meal as a protein source for supplementation of young Nellore bulls grazing in tropical pastures without affecting animal performance, NEU, MCP, and CH4 emissions.

Progressive inclusion of pearl millet herbage as a supplement for dairy cows fed mixed rations: Effects on methane emissions, dry matter intake, and milk production.

The inclusion of grazing in dairy feeding systems can improve animal welfare and reduce feed costs and labor for animal care and manure management. This work aimed to evaluate the effects of including pearl millet herbage (Pennisetum glaucum 'Campeiro') as a supplement for dairy cows fed total mixed rations (TMR). The treatments included 100% TMR offered ad libitum (control, TMR100), 75% TMR ad libitum intake + access to grazing of a pearl millet pasture between the morning and afternoon milkings (7 h/d; pTMR75), and 50% TMR ad libitum intake + access to grazing of a pearl millet pasture between the morning and afternoon milkings (7 h/d; pTMR50). Nine multiparous Holstein and F1 Jersey × Holstein cows were distributed in a replicated 3 × 3 Latin square design with 3 periods of 21 d (a 16-d adaptation period and a 5-d measurement period). Cows in the TMR75 and TMR50 groups strip-grazed a pearl millet pasture with pre- and postgrazing sward height targets of 60 and 30 cm, respectively. The herbage dry matter intake (DMI) increased with decreasing mixed ration supplies, and the total DMI decreased linearly from 19.0 kg/d in the TMR100 group to 18.0 kg/d in the pTMR50 group. Milk production decreased linearly from 24.0 kg/d in the TMR100 group to 22.4 kg/d in the pTMR50 group, and energy-corrected milk (ECM) production decreased linearly from 26.0 kg/d to 23.6 kg/d. Enteric methane (CH4) emissions decreased linearly from 540 g/d in the TMR100 group to 436 g/d in the pTMR50 group, and CH4 yields (g/kg of DMI) tended to decrease linearly. The CH4 intensity was similar between treatments, averaging 20 g of CH4/kg of ECM. The inclusion of pearl millet herbage in the dairy cow diets decreased the total DMI and milk production to a small extent without affecting CH4 intensity (g/kg of ECM).

Greenhouse gas balance and carbon footprint of pasture-based beef cattle production systems in the tropical region (Atlantic Forest biome).

The production of beef cattle in the Atlantic Forest biome mostly takes place in pastoral production systems. There are millions of hectares covered with pastures in this biome, including degraded pasture (DP), and only small area of the original Atlantic Forest has been preserved in tropics, implying that actions must be taken by the livestock sector to improve sustainability. Intensification makes it possible to produce the same amount, or more beef, in a smaller area; however, the environmental impacts must be assessed. Regarding climate change, the C dynamics is essential to define which beef cattle systems are sustainable. The objectives of this study were to investigate the C balance (t CO2e./ha per year), the intensity of C emission (kg CO2e./kg BW or carcass) and the C footprint (t CO2e./ha per year) of pasture-based beef cattle production systems, inside the farm gate and considering the inputs. The results were used to calculate the number of trees to be planted in beef cattle production systems to mitigate greenhouse gas (GHG) emissions. The GHG emission and C balance, for 2 years, were calculated based on the global warming potential (GWP) of AR4 and GWP of AR5. Forty-eight steers were allotted to four grazing systems: DP, irrigated high stocking rate pasture (IHS), rainfed high stocking rate pasture (RHS) and rainfed medium stocking rate pasture (RMS). The rainfed systems (RHS and RMS) presented the lowest C footprints (-1.22 and 0.45 t CO2e./ha per year, respectively), with C credits to RMS when using the GWP of AR4. The IHS system showed less favorable results for C footprint (-15.71 t CO2e./ha per year), but results were better when emissions were expressed in relation to the annual BW gain (-10.21 kg CO2e./kg BW) because of its higher yield. Although the DP system had an intermediate result for C footprint (-6.23 t CO2e./ha per year), the result was the worst (-30.21 CO2e./kg BW) when the index was expressed in relation to the annual BW gain, because in addition to GHG emissions from the animals in the system there were also losses in the annual rate of C sequestration. Notably, the intensification in pasture management had a land-saving effect (3.63 ha for IHS, 1.90 for RHS and 1.19 for RMS), contributing to the preservation of the tropical forest.

Host-pathogen interaction in Toxoplasma gondii-infected mixed chicken blood cell cultures.

Toxoplasma gondii has the ability to infect various nucleated cell types in different hosts. The aim of the present study was to investigate which chicken blood cells were targeted by T. gondii in a mixed blood cell culture similar to in vivo conditions and to evaluate parasite-host cell interactions. The study consisted of two subsequent experiments. In experiment 1, we applied T. gondii tachyzoites (ME49) at a multiplicity of infection of 1 tachyzoite per blood cell and examined parasite replication, cytokine, and inducible nitric oxide synthase (iNOS) mRNA expression between 1 h and 48 h post-infection (p.i.) by quantitative PCR. By using T. gondii RH-GFP tachyzoites expressing the green fluorescent protein (GFP) in experiment 2, we aimed for visualizing infected cells by confocal laser scanning microscopy (CLSM) and flow cytometric analysis at 24 h p.i. The parasite replication curve showed a massive decrease of parasite stages until 24 h p.i. followed by an approximately plateau phase. We observed mainly significantly increased iNOS mRNA expression levels in T. gondii-infected culture compared to uninfected cells. Flow cytometry and CLSM data confirmed monocytes/macrophages as main target cells for T. gondii. Moreover, different lymphocytes like B cells and cytotoxic T cells seem to be targeted to a low extent. Our findings indicate that monocytes/macrophages play a key role during T. gondii infection in chicken as host cells and triggering of immune response. To the best of our knowledge, this is the first report of a mixed chicken blood cell culture experimentally infected with T. gondii.

The effect of grazing system intensification on the growth and meat quality of beef cattle in the Brazilian Atlantic Forest biome.

This study was carried out to evaluate the effects of four levels of intensification of grazing systems: 1) degraded pasture - DP; 2) irrigated pasture with high stocking rate - IHS; 3) dryland pasture with high stocking rate - DHS; 4) dryland pasture with moderate stocking rate - DMS; on growth, muscle development and meat quality of Nellore steers (271±2.2kg of live body weight - BW; 15months old) during two consecutive periods (17 and 15months). The final BW, the average daily BW gain, the hot carcass weight and the dress percentage were greater (P<0.0001), and the ribeye area tended to be greater (P=0.085), in the intensified systems compared to the degraded system. Animals in all systems presented similar back fat. Muscle development increased with the intensification of the grazing systems and meat quality was not affected.

Salmonella Enteritidis with double deletion in phoP fliC and a competitive exclusion culture elicit substantial additive protective effects against Salmonella exposure in newly hatched chicks.

A live Salmonella Enteritidis vaccine (SE147N ΔphoP fliC), able to express both a homologous intestinal colonisation-inhibition effect and a systemic invasion-inhibition effect, was tested for its potential to generate a postulated additive protective effect in case of combined application with a competitive exclusion (CE) culture against Salmonella exposure in very young chicks. Both, SE147N ΔphoP fliC and the CE culture alone were highly protective against systemic and intestinal colonisation of the challenge strain in case of moderate Salmonella exposure, consequently, additive protective effects in combined use could not be detected. However, in case of high Salmonella Enteritidis challenge with 106 cfu/bird at day 3 of life the combination of the ΔphoP fliC vaccine and the CE culture resulted in a protective effect much more pronounced than either of the single preparations and most substantial compared to untreated control birds. The term additive protective effects reflects the recognition that exclusion effects by gut flora cultures and inhibition effects by Salmonella vaccines are caused by different mechanisms.

Prediction of the chemical body composition of Nellore and crossbreed bulls.

Young Nellore and crossbreed bulls were comparatively slaughtered to generate equation models for predicting the chemical composition of the empty body and carcass from the chemical composition of the Hankins and Howe section (; ). Data were collected from 236 animals from different genetic groups: Nellore, one-half Canchim + one-half Nellore, one-half Angus + one-half Nellore, and one-half Simmental + one-half Nellore, with 48 baseline animals (BW range from 218 to 433 kg) and 188 animals finished in the feedlot (BW range from 356 to 618 kg). The chemical composition prediction equation model was developed for all genetic groups using stepwise regression analysis. Across all animals, the percentages of water and ether extract in the HH section were highly correlated ( < 0.001) with the percentages in the carcass ( = 0.911 and = 0.901, respectively, for water content of the carcass [HOC] and = 0.921 and = 0.921, respectively, for ether extract content of the carcass [EEC]) and empty body ( = 0.937 and = 0.926, respectively, for water content of the empty body [HOEB] and = 0.935 and = 0.939, respectively, for ether extract content of the empty body [EEEB]). The best prediction models were for the traits of empty body weight, HOEB, EEEB, HOC, and EEC. Determination coefficients for predicting the dependent variables obtained from the carcass composition were lower than those obtained from the empty body composition. It was concluded that the chemical composition of the empty body and the carcass can be predicted from the composition of the HH section, using a general equation for different genetic groups.

Effect of the genetic group, production system and sex on the meat quality and sensory traits of beef from crossbred animals.

The crossbreeding of two or more breeds from the Bos taurus and Bos indicus species is an alternative for obtaining high-quality meat from animals adapted to tropical climates. Quality and sensory attributes of beef, mainly its tenderness and flavour, are very important with regard to the consumer's point of view. This study aimed to evaluate the physico-chemical and sensory characteristics of crossbred young bulls and heifers, the offspring of Angus or Limousin bulls and 1/2 Angus + 1/2 Nellore or 1/2 Simental + 1/2 Nellore cows that were finished on feedlot or pasture. Meat quality traits (pH, colour, cooking loss, water holding capacity and shear force) and sensory parameters (characteristic beef aroma/flavour intensity, strange aroma/flavour intensity, tenderness and juiciness descriptive attributes, flavour, texture (tenderness) and overall acceptance) were evaluated. The genetic group had an effect on the beef pH, but it was not as relevant as the effect of the combination between the production system and the sex or genetic group, which affected many of the quality and sensory traits.

Combination of competitive exclusion and immunisation with a live Salmonella vaccine in newly hatched chickens: Immunological and microbiological effects.

In addition to evaluating the efficacy potential of a combined use of vaccination and competitive exclusion (CE) against Salmonella exposure in chicks at 3-days of age, a live Salmonella Enteritidis vaccine (SE-LV) and a CE culture were tested for their ability to induce parameters of the innate immunity. Whereas the invasive SE-LV induced an influx of granulocytes and macrophages as well as an increased transcription of several cytokines in the caecal mucosa, the CE culture did not demonstrate any differences in these parameters compared to controls. It is therefore highly probable that the effects observed with CE cultures are not due to the rapid stimulation of the immune system. The combined use of both preparations did not result in an additive intestinal exclusion effect of the challenge strain more pronounced than that after single administration of the CE culture. The combined use of the Salmonella live vaccine and the CE culture resulted in an additive protective effect and prevented completely the systemic dissemination of the Salmonella challenge strain. To exploit the potential of combined use of CE and vaccination further and most effectively, live Salmonella vaccines are needed that are despite their attenuation in virulence still capable to induce both intestinal colonisation- and invasion-inhibition effects against Salmonella exposure.

Approaches to Investigating Complex Genetic Traits in a Large-Scale Inbred Mouse Aging Study.

Inbred mice are a unique model system for studying aging because of the genetic homogeneity within inbred strains, the short life span of mice relative to humans, and the rich array of analytic tools that are available. A large-scale aging study was conducted on 28 inbred strains representing great genetic diversity to determine, via histopathology, the type and diversity of spontaneous diseases that aging mice develop. A total of 20 885 different diagnoses were made, with an average of 12 diagnoses per mouse in the study. Eighteen inbred strains have had their genomes sequenced, and many others have been partially sequenced to provide large repositories of data on genetic variation among the strains. This vast amount of genomic information can be utilized in genome-wide association studies to find candidate genes that are involved in the pathogenesis of spontaneous diseases. As an illustration, this article presents a genome-wide association study of the genetic associations of age-related intestinal amyloidosis, which implicated 3 candidate genes: translocating chain-associated membrane protein 1 (Tram1); splicing factor 3b, subunit 5 (Sf3b5); and syntaxin 11 (Stx11). Representative photomicrographs are available on the Mouse Tumor Biology Database and Pathbase to serve as a reference when evaluating inbred mice used in other genetic or experimental studies to rule out strain background lesions. Many of the age-related mouse diseases are similar, if not identical, to human diseases; therefore, the genetic discoveries have direct translational benefit.

Effect of lipid sources with different fatty acid profiles on the intake, performance, and methane emissions of feedlot Nellore steers.

The present study was conducted to evaluate the effect of lipids with different fatty acid profiles on the intake, performance, and enteric CH4 emission of Nellore steers. A total of 45 Nellore animals with an average initial BW of 419 ± 11 kg (at 15 ± 2 mo) were distributed in a completely randomized design consisting of 5 treatments and 9 replicates. The roughage feed was maize silage (600 g/kg on a DM basis) plus concentrate (400 g/kg on a DM basis). The dietary treatments were as follows: without additional fat (WF), palm oil (PO), linseed oil (LO), protected fat (PF; Lactoplus), and whole soybeans (WS). The lipid source significantly affected (P < 0.05) nutrient intake. The greatest intakes of DM, OM, and CP were observed in the animals that were fed the WF or PF diets, and the lowest intakes were observed in the animals that were fed the PO diet. Intake of NDF decreased (P < 0.05) with the addition of PO. Enteric methane emission (g/kg DMI) was reduced by an average of 30% when the animals were fed diets containing WS, LO, and PO (P < 0.05), and these diets caused a larger reduction in the energy loss in the form of methane compared to those without added fat and with added PF (3.3 vs. 4.7%). The different fatty acid profiles did not affect the backfat thickness or the loin eye area of the animals (P > 0.05). However, animals fed PO displayed lower daily weight gain (0.36 kg/d), feed efficiency (0.08 kg ADG/kg DM), HCW (245 kg), and hot yield percentage (52.6%) compared to animals that were fed the other diets. Therefore, PO compared to the other lipid sources used in this study reduces intake, performance, feed efficiency, and carcass yield. Therefore, PO is not suggested for feedlot-finished animals.

Phenotypic characterization of the KK/HlJ inbred mouse strain.

Detailed histopathological diagnoses of inbred mouse strains are important for interpreting research results and defining novel models of human diseases. The aim of this study was to histologically detect lesions affecting the KK/HlJ inbred strain. Mice were examined at 6, 12, and 20 months of age and near natural death (ie, moribund mice). Histopathological lesions were quantified by percentage of affected mice per age group and sex. Predominant lesions were mineralization, hyperplasia, and fibro-osseous lesions. Mineralization was most frequently found in the connective tissue dermal sheath of vibrissae, the heart, and the lung. Mineralization was also found in many other organs but to a lesser degree. Hyperplasia was found most commonly in the pancreatic islets, and fibro-osseous lesions were observed in several bones. The percentage of lesions increased with age until 20 months. This study shows that KK/HlJ mice demonstrate systemic aberrant mineralization, with greatest frequency in aged mice. The detailed information about histopathological lesions in the inbred strain KK/HlJ can help investigators to choose the right model and correctly interpret the experimental results.

Energy efficiency of growing ram lambs fed concentrate-based diets with different roughage sources.

Poor-quality roughages are widely used as fiber sources in concentrate-based diets for ruminants. Because roughage quality is associated with the efficiency of energy use in forage-based diets, the objective of this study was to determine whether differing the roughage source in concentrate-based diets could change the energy requirements of growing lambs. Eighty-four 1/2 Dorper × 1/2 Santa Inês ram lambs (18.0 ± 3.3 kg BW) were individually penned and divided into 2 groups according to primary source of dietary roughage: low-quality roughage (LQR; sugarcane bagasse) or medium-quality roughage (MQR; coastcross hay). Diets were formulated to be isonitrogenous (2.6% N) and to meet 20% of physically effective NDF. After a 10-d ad libitum adaptation period, 7 lambs from each group were randomly selected and slaughtered (baseline). Twenty-one lambs in each diet group were fed ad libitum and slaughtered at 25, 35, or 45 kg BW. The remaining 28 lambs (14 from each diet group) were submitted to 1 of 2 levels of feed restriction: 70% or 50% of the ad libitum intake. Retentions of body fat, N, and energy were determined. Additionally, 6 ram lambs (44.3 ± 5.6 kg BW) were kept in metabolic cages and used in a 6 × 6 Latin square experiment designed to establish the ME content of the 2 diets at the 3 levels of DM intake. There was no effect of intake level on diet ME content, but it was greater in the diet with LQR than in the diet with MQR (3.18 vs. 2.94 Mcal/kg, respectively; P < 0.01). Lambs fed the diet with LQR had greater body fat (g/kg of empty BW) and energy concentrations (kcal/kg of empty BW) because of a larger visceral fat deposition (P < 0.05). Using a low-quality roughage as a primary source of forage in a concentrate-based diet for growing lambs did not change NEm and the efficiency of ME use for maintenance, which averaged 71.6 kcal/kg(0.75) of shrunk BW and 0.63, respectively. On the other hand, the greater nonfibrous carbohydrate content of the diet with LQR resulted in a 17% better efficiency of ME use for gain (P < 0.01), which was associated with a greater partial efficiency of energy retention as fat (P < 0.01). This increased nutritional efficiency, however, should be viewed with caution because it is related to visceral fat deposition, a nonedible tissue.

Measurement and mitigation of methane emissions from beef cattle in tropical grazing systems: a perspective from Australia and Brazil.

The growing global demand for food of animal origin will be the incentive for countries such as Australia and Brazil to increase their beef production and international exports. This increased supply of beef is expected to occur primarily through on-farm productivity increases. The strategies for reducing resultant greenhouse gas (GHG) emissions should be evaluated in the context of the production system and should encompass a broader analysis, which would include the emissions of methane (CH4) and nitrous oxide (N2O) and carbon sequestration. This paper provides an insight into CH4 measurement techniques applicable to grazing environments and proposed mitigation strategies, with relevance to the production systems that are predominant in grazing systems of Australia and Brazil. Research and technology investment in both Australia and Brazil is aimed at developing measurement techniques and increasing the efficiency of cattle production by improving herd genetics, utilization of the seasonal feed-base and reducing the proportion of metabolizable energy lost as CH4. Concerted efforts in these areas can be expected to reduce the number of unproductive animals, reduce age at slaughter and inevitably reduce emission intensity (EI) from beef production systems. Improving efficiency of livestock production systems in tropical grazing systems for Australia and Brazil will be based on cultivated and existing native pastures and the use of additives and by-products from other agricultural sectors. This approach spares grain-based feed reserves typically used for human consumption, but potentially incurs a heavier EI than current intensive feeding systems. The determination of GHG emissions and the value of mitigation outcomes for entire beef production systems in the extensive grazing systems is complex and require a multidisciplinary approach. It is fortunate that governments in both Australia and Brazil are supporting ongoing research activities. Nevertheless, to achieve an outcome that feeds a growing population while reducing emissions on a global scale continues to be a monumental challenge for ruminant nutritionists.

Phosphodiesterase 2 inhibitors promote axonal outgrowth in organotypic slice co-cultures.

The development of appropriate models assessing the potential of substances for regeneration of neuronal circuits is of great importance. Here, we present procedures to analyze effects of substances on fiber outgrowth based on organotypic slice co-cultures of the nigrostriatal dopaminergic system in combination with biocytin tracing and tyrosine hydroxylase labeling and subsequent automated image quantification. Selected phosphodiesterase inhibitors (PDE-Is) were studied to identify their potential growth-promoting capacities. Immunohistochemical methods were used to visualize developing fibers in the border region between ventral tegmental area/substantia nigra co-cultivated with the striatum as well as the cellular expression of PDE2A and PDE10. The quantification shows a significant increase of fiber density in the border region induced by PDE2-Is (BAY60-7550; ND7001), comparable with the potential of the nerve growth factor and in contrast to PDE10-I (MP-10). Analysis of tyrosine hydroxylase-positive fibers indicated a significant increase after treatment with BAY60-7550 and nerve growth factor in relation to dimethyl sulfoxide. Additionally, a dose-dependent increase of intracellular cGMP levels in response to the applied PDE2-Is in PDE2-transfected HEK293 cells was found. In summary, our findings show that PDE2-Is are able to significantly promote axonal outgrowth in organotypic slice co-cultures, which are a suitable model to assess growth-related effects in neuro(re)generation.

Dietary nitrate supplementation reduces methane emission in beef cattle fed sugarcane-based diets.

The objective of this study was to determine the effect of dietary nitrate on methane emission and rumen fermentation parameters in Nellore × Guzera (Bos indicus) beef cattle fed a sugarcane based diet. The experiment was conducted with 16 steers weighing 283 ± 49 kg (mean ± SD), 6 rumen cannulated and 10 intact steers, in a cross-over design. The animals were blocked according to BW and presence or absence of rumen cannula and randomly allocated to either the nitrate diet (22 g nitrate/kg DM) or the control diet made isonitrogenous by the addition of urea. The diets consisted of freshly chopped sugarcane and concentrate (60:40 on DM basis), fed as a mixed ration. A 16-d adaptation period was used to allow the rumen microbes to adapt to dietary nitrate. Methane emission was measured using the sulfur hexafluoride tracer technique. Dry matter intake (P = 0.09) tended to be less when nitrate was present in the diet compared with the control, 6.60 and 7.05 kg/d DMI, respectively. The daily methane production was reduced (P < 0.01) by 32% when steers were fed the nitrate diet (85 g/d) compared with the urea diet (125 g/d). Methane emission per kilogram DMI was 27% less (P < 0.01) on the nitrate diet (13.3 g methane/kg DMI) than on the control diet (18.2 g methane/kg DMI). Methane losses as a fraction of gross energy intake (GEI) were less (P < 0.01) on the nitrate diet (4.2% of GEI) than on the control diet (5.9% of GEI). Nitrate mitigated enteric methane production by 87% of the theoretical potential. The rumen fluid ammonia-nitrogen (NH(3)-N()) concentration was significantly greater (P < 0.05) for the nitrate diet. The total concentration of VFA was not affected (P = 0.61) by nitrate in the diet, while the proportion of acetic acid tended to be greater (P = 0.09), propionic acid less (P = 0.06) and acetate/propionate ratio tended to be greater (P = 0.06) for the nitrate diet. Dietary nitrate reduced enteric methane emission in beef cattle fed sugarcane based diet.

Emergency and therapeutic vaccination--is stimulating innate immunity an option?

There is increasing evidence that activation of innate immunity, in animals and man, by live vaccines, sub-unit vaccines or synthetic or non-synthetic stimulants can induce a profound and rapidly induced resistance to pathogens, including infectious agents that are unrelated to the stimulating antigen or agent. We review the evidence for this phenomenon and present the proposition that this approach might be used to stimulate immunity during the life of the animal when susceptibility to infection is high and when normal vaccination procedures may be inappropriate.

Regulation of lipid binding underlies the activation mechanism of class IA PI3-kinases.

Somatic missense mutations in PIK3CA, which encodes the p110α catalytic subunit of phosphoinositide 3-kinases, occur frequently in human cancers. Activating mutations spread across multiple domains, some of which are located at inhibitory contact sites formed with the regulatory subunit p85α. PIK3R1, which encodes p85α, also has activating somatic mutations. We find a strong correlation between lipid kinase and lipid-binding activities for both wild-type (WT) and a representative set of oncogenic mutant complexes of p110α/p85α. Lipid binding involves both electrostatic and hydrophobic interactions. Activation caused by a phosphorylated receptor tyrosine kinase (RTK) peptide binding to the p85α N-terminal SH2 domain (nSH2) induces lipid binding. This depends on the polybasic activation loop as well as a conserved hydrophobic motif in the C-terminal region of the kinase domain. The hotspot E545K mutant largely mimics the activated WT p110α. It shows the highest basal activity and lipid binding, and is not significantly activated by an RTK phosphopeptide. Both the hotspot H1047R mutant and rare mutations (C420R, M1043I, H1047L, G1049R and p85α-N564D) also show increased basal kinase activities and lipid binding. However, their activities are further enhanced by an RTK phosphopeptide to levels markedly exceeding that of activated WT p110α. Phosphopeptide binding to p110ß/p85α and p110δ/p85α complexes also induces their lipid binding. We present a crystal structure of WT p110α complexed with the p85α inter-SH2 domain and the inhibitor PIK-108. Additional to the ATP-binding pocket, an unexpected, second PIK-108 binding site is observed in the kinase C-lobe. We show a global conformational change in p110α consistent with allosteric regulation of the kinase domain by nSH2. These findings broaden our understanding of the differential biological outputs exhibited by distinct types of mutations regarding growth factor dependence, and suggest a two-tier classification scheme relating p110α and p85α mutations with signalling potential.