Peripheral and gastrointestinal immune systems of healthy cattle raised outdoors at pasture or indoors on a concentrate-based ration

Background  

Despite an increasing preference of consumers for beef produced from more extensive pasture-based production systems and potential human health benefits from the consumption of such beef, data regarding the health status of animals raised on pasture are limited. The objective of this study was to characterise specific aspects of the bovine peripheral and the gastrointestinal muscosal immune systems of cattle raised on an outdoor pasture system in comparison to animals raised on a conventional intensive indoor concentrate-based system.     

The effects of graded levels of concentrate supplementation on colour and lipid stability of beef from pasture finished late-maturing bulls

Finishing late-maturing bulls on grass may alter the antioxidant/prooxidant balance leading to beef with higher susceptibility to lipid oxidation and a lower colour stability compared to bulls finished on cereal concentrates. In this context, lipid oxidation and colour stability of beef from late-maturing bulls finished on pasture, with or without concentrate supplements, or indoors on concentrate was assessed. Charolais or Limousin sired bulls (n = 48) were assigned to four production systems: (1) pasture only (P), (2) pasture plus 25% dietary DM intake as barley-based concentrate (PC25), (3) pasture plus 50% dietary DM intake as barley-based concentrate (PC50) or (4) a barley-based concentrate ration (C). Following slaughter and postmortem ageing, M. Longissimus thoracis et lumborum was subjected to simulated retail display (4°C, 1000 lux for 12 h out of 24 h) for 3, 7, 10 and 14 days in modified atmosphere packs (O₂ : CO₂; 80 : 20). Lipid oxidation was determined using the 2-thiobarbituric acid-reactive substances assay; α-tocopherol was determined by HPLC; fatty acid methyl esters were determined using Gas Chromatography. Using a randomised complete block design, treatment means were compared by either ANOVA or repeated measures ANOVA using the MIXED procedure of SAS. Total polyunsaturated fatty acid (PUFA) concentrations were not affected by treatment, n-3 PUFAs were higher (P < 0.001) and the ratio of n-6 to n-3 PUFAs was lower (P < 0.001) in muscle from P, PC25 and PC50 compared to C. α-Tocopherol concentration was higher in muscle from P compared to PC50 and C bulls (P = 0.001) and decreased (P < 0.001) in all samples by day 14. Lipid oxidation was higher in muscle from C compared to P bulls on day 10 and day 14 of storage (P < 0.01). Finishing on pasture without supplementation did not affect beef colour stability and led to lower lipid oxidation, possibly due to the higher α-tocopherol concentration compared to concentrate finished beef.     

Water footprinting of pasture-based farms; beef and sheep

In the context of water use for agricultural production, water footprints (WFs) have become an important sustainability indicator. To understand better the water demand for beef and sheep meat produced on pasture-based systems, a WF of individual farms is required. The main objective of this study was to determine the primary contributors to freshwater consumption up to the farm gate expressed as a volumetric WF and associated impacts for the production of 1 kg of beef and 1 kg of sheep meat from a selection of pasture-based farms for 2 consecutive years, 2014 and 2015. The WF included green water, from the consumption of soil moisture due to evapotranspiration, and blue water, from the consumption of ground and surface waters. The impact of freshwater consumption on global water stress from the production of beef and sheep meat in Ireland was also computed. The average WF of the beef farms was 8391 l/kg carcass weight (CW) of which 8222 l/kg CW was green water and 169 l/kg CW was blue water; water for the production of pasture (including silage and grass) contributed 88% to the WF, concentrate production – 10% and on-farm water use – 1%. The average stress-weighted WF of beef was 91 l H₂O eq/kg CW, implying that each kg of beef produced in Ireland contributed to freshwater scarcity equivalent to the consumption of 91 l of freshwater by an average world citizen. The average WF of the sheep farms was 7672 l/kg CW of which 7635 l/kg CW was green water and 37 l/kg CW was blue water; water for the production of pasture contributed 87% to the WF, concentrate production – 12% and on-farm water use – 1%. The average stress-weighted WF was 2 l H₂O eq/kg CW for sheep. This study also evaluated the sustainability of recent intensification initiatives in Ireland and found that increases in productivity were supported through an increase in green water use and higher grass yields per hectare on both beef and sheep farms.     

Restricting dairy cow access time to pasture in early lactation: the effects on milk production, grazing behaviour and dry matter intake

One of the main aims of pasture-based systems of dairy production is to increase the proportion of grazed grass in the diet. This is most easily achieved by increasing the number of grazing days. However, periods of inclement weather conditions can reduce the number of days at pasture. The two objectives of this experiment were: (i) to investigate the effect of restricting pasture access time on animal production, grazing behaviour and dry matter intake (DMI) of spring calving dairy cows in early lactation; and (ii) to establish whether silage supplementation is required when cows return indoors after short grazing periods. In all, 52 Holstein-Friesian spring calving dairy cows were assigned to a four-treatment study from 25 February to 26 March 2008. The four treatments were: full-time access to pasture (22H; control); 4.5-h- pasture access after both milkings (2 × 4.5H); 3-h pasture access after both milkings (2 × 3H); 3-h pasture access after both milkings with silage supplementation by night (2 × 3SH). All treatments were offered 14.4 kg DM/cow per day herbage from swards, with a mean pre-grazing yield of 1739 kg DM/ha above 4 cm, - and were supplemented with 3 kg DM/cow per day of concentrate. The 2 × 3SH treatment was offered an additional 4 kg DM/cow of grass silage by night. Restricting pasture access time (2 × 3H, 2 × 3SH and 2 × 4.5H) had no effect on milk (28.3 kg/cow per day) and solids-corrected milk (27.2 kg/cow per day) yield when compared with the treatment grazing full time. Supplementing animals with grass silage did not increase milk production when compared with all other treatments. Milk protein concentration tended to be lower (P = 0.08; 32.2 g/kg) for the 2 × 3SH animals when compared with the 22H animals (33.7 g/kg). The grass DMI of the 2 × 3SH treatment was significantly lower (-2.3 kg DM/cow per day) than all other treatments (11.9 kg DM/cow per day), yet the total DMI of these animals was highest (16.6 kg DM/cow per day). The 22H cows grazed for 481 min/cow per day, which is significantly longer than all other treatments. The 2 × 3H animals grazed for 98% of the time, whereas the 2 × 3SH grazed for 79% of their time at pasture. Restricting pasture access time did not affect end body weight or body condition score. The results of this study indicate that restricting pasture access time of dairy cows in early lactation does not affect milk production performance. Furthermore, supplementing cows with grass silage does not increase milk production but reduces grazing efficiency.     

Growth performance,carcass trait,meat quality and oxidative stability of beef cattle offered alternative silages in a finishing ration

As lack of forage resource, alternative roughage sources have been developed for ruminant production and their inclusion would exert a great effect on the dietary nutrition, consequently affecting animal performance. Four silages (corn silage (CS), corn stalk silage (SS), inoculated CS and inoculated SS) were separately offered to 60 Bohai Black cattle (15 cattle/group) during a 24-week finishing period, in which the growth performance, carcass trait, beef quality and oxidative stability of steers were determined. Neither silage material nor silage inoculant exerted a significant effect on the growth performance, carcass trait and oxidative stability of beef cattle (P>0.05). As to beef quality, cattle offered CS had higher (P<0.05) contents of intramuscular fat than those offered SS along with a lower moisture content (P<0.05). The contents (mg/g muscle) of C10 : 0, C12 : 0, C14 : 1, C16 : 0, C16 : 1, C18 : 1n9c, C18 : 2n6c, C18 : 3n3, C20 : 1n9, C20 : 2, C20 : 3n6, saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids and n-6 fatty acids were higher (P<0.05) in the beef muscle of animals offered CS than those offered SS, whereas inoculated treatment made no difference (P>0.05) on the proximate components and fatty acids profile of beef muscle. There was neither an interaction (P>0.05) between inoculated treatment and silage material. There were no differences (P>0.05) in cholesterol content and meat quality traits in animals fed alternative silages. The collective findings suggest that it is not economical to substitute high-quality forage for relative low-quality forage in a high-concentrate finishing ration of beef cattle and silage inoculant inclusion would not exert a direct effect on animal performance.     

Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems

We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The μ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.     

Evaluation of concentrate factors affecting silage intake of dairy cows: a development of the relative total diet intake index

The aim of this work was to develop an index describing the relative intake of the total diet by dairy cows, and hence the ability to predict intake responses to changes in both forage and concentrate variables. An evaluation of concentrate factors affecting silage dry matter (DM) intake of dairy cows was conducted based on dietary treatment means from milk production experiments. The data were divided into four subsets according to concentrate treatments used within the experiments: the amount of concentrate supplementation (n = 217), protein supplementation (n = 336), carbohydrate composition (n = 114) and fat concentration of the concentrate (n = 29). The data were subjected to mixed-model regression analysis. Increased concentrate DM intake (CDMI) decreased silage DM intake (SDMI) quadratically. The substitution rate (substitution of silage DM for concentrate DM) increased with improved silage intake potential. SDMI increased quadratically with concentrate protein intake, the response being negatively related to the effective protein degradability (EPD) of concentrates. Replacement of starchy concentrate ingredients with fibrous supplements had a small positive effect on silage intake, whereas increased concentrate fat concentration slightly decreased SDMI. The outcome of concentrate factors influencing total DM intake (TDMI) was used to create a relative CDMI index as follows: CDMI index = 100 + 10 × [(CDMI - 0.1629 × CDMI - 0.01882 × CDMI2 - 5.49) + ((0.9474 × CCPI - 0.4965 × CCPI2) - 2.017 × (CEPD - 0.74)) + 0.00225 × (CNDF - 250) - 0.0103 × (40 - Cfat) - 0.00058 × (CDMI - 8.0) × (SDMI index - 100)], where CDMI = concentrate DM intake (kg/day), CCPI = supplementary concentrate CP intake (kg/day; CP>170 g/kg DM), CEPD = concentrate EPD (g/g), CNDF = concentrate NDF concentration (g/kg DM), Cfat = concentrate fat concentration (g/kg DM) and SDMI index is the relative intake potential of silage (Huhtanen, Rinne and Nousiainen 2007. Animal 1, 758-770). TDMI index was calculated as SDMI index + CDMI index - 100 to describe the relative intake potential of the total diet. For the whole data set (n = 943), one TDMI index unit was equivalent to 0.095 kg/day DM intake, i.e. close to the default value of 0.100 kg. The CDMI index explained proportionally 0.88 of the variation in TDMI within a study with a 0.27 kg/day residual mean-square error (n = 616). The corresponding values for the TDMI index were 0.81 and 0.37 kg/day (n = 943), respectively. The residual mean-square errors in cross-validation were marginally higher. The developed TDMI index can be used to estimate the intake responses to diet changes. It provides an improved basis for practical dairy cow ration formulation and economic evaluation.     

Comparison of organoleptic quality and composition of beef from suckler bulls from different production systems

Bull beef production is traditionally based on high concentrate rations fed indoors. Inclusion of grazed grass, which is generally a cheaper feed, would decrease the cost of bull beef production, but may affect beef quality. Accordingly, the organoleptic quality and composition of beef from continental-sired suckler bulls (n=126) assigned to either ad libitum concentrates to slaughter (C), grass silage (GS) ad libitum for 120 days followed by C (GSC) or GS followed by 100 days at pasture and then C (GSPC) and slaughtered at target carcass weights (CW) of 360, 410 or 460 kg was examined. Tenderness, flavour liking and overall liking were lower (P<0.05) for GSPC than for C and GSC. Intramuscular fat content and soluble collagen proportion were lower (P<0.05) for GSPC than GSC which was lower (P<0.05) than C. Soluble collagen proportion was lower (P<0.05) for 460 kg than 410 kg CW, which was lower (P<0.05) than 360 kg CW. Inclusion of a grazing period decreased the ratings of tenderness, flavour liking and overall liking, but age of the bulls at slaughter had no clear influence on sensory characteristics.     

Consequential Life Cycle Assessment of Pasture‐based Milk Production: A Case Study in the Waikato Region,New Zealand

Farm intensification options in pasture‐based dairy systems are generally associated with increased stocking rates coupled with the increased use of off‐farm inputs to support the additional feed demand of animals. However, as well as increasing milk production per hectare, intensification can also exacerbate adverse impacts on the environment. The objective of the present study was to investigate environmental trade‐offs associated with potential intensification methods for pasture‐based dairy farming systems in the Waikato region, New Zealand. The intensification scenarios selected were (1) increased pasture utilization efficiency (PUE scenario), (2) increased use of nitrogen (N) fertilizer to boost on‐farm pasture production (N fertilizer scenario), and (3) increased use of brought‐in feed as maize silage (MS) (MS scenario). Twelve impact categories were assessed. The PUE scenario was the environmentally preferred intensification method, and the preferred choice between the N fertilizer and MS scenarios depended upon trade‐offs between different environmental impacts. Sensitivity analysis was carried out to test the effects of choice associated with: (1) the approaches used to account for indirect land‐use change (ILUC) and (2) the competing product systems (conventional beef systems) used to handle the co‐product dairy meat for the climate change (CC) indicator. Results showed that the magnitude of the CC indicator results was influenced by the ILUC accounting approaches and the choice associated with a global marginal beef mix, but the relative CC indicator results for the three intensification scenarios remained unchanged.     

Length of concentrate finishing affects the fatty acid composition of grass-fed and genetically lean beef: an emphasis on trans-18:1 and conjugated linoleic acid profiles

Intensively finishing cattle on a high-grain diet is generally used to enhance marbling, whereas extensively finishing on grass is known to provide improved muscle fatty acid profiles. The objective of this study was to evaluate to what extent intensive concentrate finishing (0, 1 or 2 months) can be combined with forage feeding without negatively affecting the fatty acid profile of genetically lean animals. Bulls from the 'Asturiana de los Valles' breed were reared under grazing conditions with/without final finishing on a barley-based concentrate: 0 months (control; n=7), 1 month (n=10) and 2 months (n=7). Yearling bulls were slaughtered commercially at an average live weight of 516±9.8 kg. Increasing the finishing time on concentrate significantly increased the saturated and monounsaturated fatty acids, whereas polyunsaturated fatty acids (PUFAs) tended to decrease and it was not possible to increase the long-chain PUFA content in muscle tissue of this breed. An increase was observed for total trans-18:1 (average 5.5% with grain v. 3.7% for grass). The 11t-18:1/10t-18:1 ratio was significantly higher in grass-fed (average 8.1) compared with grain-finished animals (average 1.1). Grass or limited concentrate finishing reduced the n-6/n-3 ratio in muscle tissue (average 3.6 for 0 and 1 month, and 4.9 for 2 months on grain finishing). The beef was within or close to the recommended values for human consumption (i.e. polyunsaturated/saturated>0.45, n-6/n-3<4.0), and total trans-FA content was low. However, finishing increased the content of undesirable trans-18:1 and conjugated linoleic acid isomers, particularly after 2 months, whereas grass finishing was judged to provide a healthier beef fatty acid profile.     

Beef palatability and its relationship with protein degradation and muscle fibre type profile in longissimus thoracis in Alentejana breed from divergent growth pathways

The traditional beef production in the South of Portugal is based on a discontinuous growth (DG) system that requires lower external inputs and could enhance meat quality and financial returns to cattle producers. This system allows farmers to take advantage of the bull’s compensatory growth when the pasture is abundant and finishes the cattle on concentrates for 2 to 3 months before slaughter. The fast gain rate before slaughter could be a valuable strategy to improve tenderness and to reduce its inconsistency in beef production. Therefore, the aim of this study was to evaluate the effects of production system (continuous growth (CG) v. DG) on longissimus thoracis muscle properties from Alentejana bulls. In total, 40 Alentejana male calves were allocated to two distinct feeding regimes: in the CG system, animals were fed concentrate plus hay and slaughtered at 18 months of age, whereas in the DG system, animals were fed on hay until 15 months of age and then fed the same diet provided to the CG group until 24 months of age. The DG system had a positive impact on meat tenderness (P<0.001) and global acceptability (P<0.001). DG bulls had greater fibre cross-sectional area (CSA) of glycolytic fibres (P<0.05) and relative area of the muscle (RA) occupied by type IIX fibres (P<0.01) and greater levels of α-actinin (P<0.05) and myosin light chain 2 (P<0.01) proteins, and pH_24h (P<0.01) than CG bulls. The latter had greater CSA of type I (P<0.05) and type IIA (P<0.01) and greater RA of type IIA (P<0.05) and oxidative (P<0.05) than CG bulls. The compensatory growth production system had a positive impact on meat tenderness and global acceptability, overcoming the negative effects of slaughter of the bulls at a later age. The DG beef system could be a worthwhile strategy of beef production in Mediterranean areas due to the low-quality pasture in summer.     

Performance,profitability and greenhouse gas emissions of alternative finishing strategies for Holstein-Friesian bulls and steers

Modifying finishing strategies within established production systems has the potential to increase beef output and farm profit while reducing greenhouse gas (GHG) emissions. Thus, the objectives of this study were to investigate the effects of finishing duration on animal performance of Holstein-Friesian (HF) bulls and steers and evaluate the profitability and GHG emissions of these finishing strategies. A total of 90 HF calves were assigned to a complete randomised block design; three bull and three steer finishing strategies. Calves were rotationally grazed in a paddock system for the first season at pasture, housed and offered grass silage ad libitum plus 1.5 kg DM of concentrate per head daily for the first winter and returned to pasture for a second season. Bulls were slaughtered at 19 months of age and either finished indoors on concentrates ad libitum for 100 days (19AL), finished at pasture supplemented with 5 kg DM of concentrate per head daily for 100 (19SP) or 150 days (19LP). Steers were slaughtered at 21 months of age and finished at pasture, supplemented with 5 kg DM of concentrate per head daily for 60 (21SP) and 110 days (21LP) or slaughtered at 24 months of age and finished indoors over the second winter on grass silage ad libitum plus 5 kg DM of concentrate per head daily (24MO). The Grange Dairy Beef Systems Model and the Beef Systems Greenhouse Gas Emissions Model were used to evaluate profitability and GHG emissions, respectively. Average daily gain during the finishing period (P<0.001), live weight at slaughter (P<0.01), carcass weight (P<0.05) and fat score (P<0.001) were greater for 19AL than 19SP and 19LP, respectively. Similarly, concentrate dry matter intake was greater for 19AL than 19SP; 19LP was intermediate (P<0.001). Live weight at slaughter (P<0.001), carcass weight (P<0.001), conformation score (P<0.05) and fat score (P<0.001) were greater for 24MO than 21SP and 21LP, respectively. During the finishing period concentrate dry matter intake was greater for 21LP than 21SP with 24MO intermediate; 542, 283 and 436 kg DM, respectively. Although pasture-based finishing strategies had lower gross output values, concentrate feed costs were also reduced thus net margin was greater than indoor finishing strategies. Reducing concentrate input increased GHG emissions for bulls and steers slaughtered at the same age, respectively. Although prolonging the finishing duration reduced GHG emissions for bull and steer production systems, finishing bulls and steers over a longer period at pasture did not enhance animal performance and profit.     

Seroprevalence of Neospora caninum infection in dairy and beef cattle in Spain

In recent years, neosporosis has been identified as a major cause of abortion in dairy and beef cattle. Although the disease has been described worldwide, there is a Jack of information concerning the prevalence of this infection in different cattle production systems. The aim of this study was to investigate the seroprevalence of Neospora caninum infection in a representative area of beef and dairy cattle production in Spain. A cross-sectional study was undertaken in which herds constituted the initial sampling unit and two strata (dairy and beef herds) were considered. Using a 95% level of confidence and setting 5% (beef) and 5.4% (dairy) error limits, 216 beef and 143 dairy herds were randomly selected and sampled. Nine animals (> 1 year old) were randomly sampled in each herd to detect the presence of the infection. A herd was considered infected when at least one animal was seropositive. In total, serum samples from 1121 dairy and 1712 beef animals were collected and tested for specific anti-N. caninum IgG using an ELISA. Specific antibodies were detected in 55.1% (119/216) beef and 83.2% (119/143) dairy herds. Individual prevalences obtained were 17.9% (306/1712) for beef and 35.9% (402/1121) for dairy animals. Presence of N. caninum infection was higher in dairy than in beef herds and the association between infection and the cattle production system (dairy or beef) was statistically significant [(chi2)Y= 29.21, P < 0.001, OR = 4.04 (2.35-6.99)]. Herd size of dairy cattle did not appear to be associated with N. caninum infection. On the contrary, infection was associated with herd size in beef cattle (chi2 = 12.79, P < 0.01). Finally, no association was found between replacement or pasture management and infection in beef herds.     

Nitrogen and energy losses and methane emissions from beef cattle fed diets with gradual replacement of maize silage and concentrate with grass silage and corn-cob mix

Diets based on large proportions of grassland-based feed are uncommon in forage-based intensive beef production, thus contradicting governmental or commercial strategies to promote the use of grassland-based feed in ruminant production systems. Compared with typical maize silage/concentrate diets, grassland-based diets are associated with impaired nitrogen (N) and energy utilisation because of the comparably lower energy and higher CP content of these feeds. However, quantitative studies concerning the effects of increased dietary proportions of grassland-derived feeds on N and energy losses and utilisation and on methane emissions are missing and the compensation potential of using a limited proportion of an energy-rich forage is unknown. Therefore, we tested five diets with varying types and proportions of forage and concentrate. Three diets consisted of grass silage, maize silage, and concentrate in ratios of, g/kg DM, 100:600:300 (G100; control), 300:500:200 (G300), and 500:300:200 (G500), respectively. Two diets were composed of grass silage, corn-cob mix (CCM), and concentrate in ratios of, g/kg DM, 500:300:200 (G500_CCM), and 750:150:100 (G750_CCM), respectively. A high-protein concentrate (270 g CP/kg DM) was fed to G100, whereas a low-protein concentrate (140 g CP/kg DM) was used in the remaining diets. Diets were fed throughout the entire fattening period to groups of six Limousin-crossbred bulls each. When weighing 246 ± 18 kg, each animal underwent a 7-day total daily faeces and urine collection, which was followed by measuring methane emissions in respiration chambers for 48 h. Total DM intake was similar across all diets, whereas the N intake varied (P < 0.05). Urinary N loss (g/day) was the highest for G750_CCM (28.2) and G100 (26.6) and lowest for G500_CCM (15.2) and G300 (16.9) (P < 0.001). Energy utilisation was comparable among all groups. Metabolisable energy intake decreased numerically only with increasing proportions of grass silage in the diet. Substituting maize silage with CCM counteracted the loss in metabolisable energy intake. Absolute methane emissions were not different across the groups, but methane emission intensity (mg/g body protein retention) varied (P < 0.05), being numerically lower for G100 (349) and G500_CCM (401) compared with the other groups (488 on average). In conclusion, the results show that the grass silage proportion in beef cattle diets can be substantially increased when strategically combined with energy-dense forages, such as CCM. This also limits the need for concentrate and additional protein sources; in addition, the associated urinary N emissions, which are potentially noxious to the environment, are avoided.     

Urinary metabolomic phenotyping of nickel induced acute toxicity in rat: an NMR spectroscopy approach

The metabolomic approach has been widely used in toxicology to investigate mechanisms of toxicity. To understand the mammalian system??s response to nickel exposure, we analysed the NiCl₂ induced metabolomic changes in urine of rats using ¹H nuclear magnetic resonance (¹H NMR) spectroscopy together with clinically relevant biochemical parameters. Male Sprague?CDawley rats were administered intraperitoneally with NiCl₂ at doses of 4, 10 and 20?mg/kg body weight. Urine samples were collected at 8, 16, 24, 72, 96 and 120?h post treatment. The metabolomic profile of rat urine showed prominent changes in citrate, dimethylamine, creatinine, choline, trimethylamine oxide (TMAO), phenyl alanine and hippurate at all doses. Principal component analysis of urine ¹H NMR spectra demonstrated the dose and time dependent development of toxicity. The metabolomic time trajectory, based on pattern recognition analysis of ¹H NMR spectra of urine, illustrated clear separation of pre and post treatments (temporal). Only animals treated with a low dose of NiCl₂ returned to normal physiology. The ¹H NMR spectral data correlated well with the clinically relevant nephrotoxic biomarkers. The urinary metabolomic phenotyping for NiCl₂ induced nephrotoxicity was defined according to the predictive ability of the known metabolite biomarkers, creatinine, citrate and TMAO. The current approach demonstrates that metabolomics, one of the most important platform in system biology, may be a promising tool for identifying and characterizing biochemical responses to toxicity.     

DNA-Based Individual and Sex Identification from Wolverine (Gulo Gulo) Faeces and Urine

Non-invasive genetic analyses are important for studies of species that are rare, sensitive or at risk of extinction. This study investigates the possibility of using faeces and urine to obtain microsatellite genotypes for individual identification of wolverines (Gulo gulo). The reliability of the employed method was assessed by analysing independent amplifications of non-invasive samples (a multiple-tube approach) and by comparing genotypes obtained from faeces to genotypes obtained from blood or tissue of the same individual. Ten microsatellite markers were successfully amplified in 65% of the faecal samples (n = 32) and 40% of the urine samples (n = 22). Allelic dropout was found in 12 and 14% of the amplifications from extracts of faeces and urine, respectively. Nevertheless, all multi-locus genotypes were correct, as judged from comparison to data from tissue or blood samples, after three replicates. These results suggest that a non-invasive approach based on DNA-analysis of faeces can be a powerful tool in population monitoring of wolverines, potentially providing reliable estimates of population size and immigration rate. A second objective of the study was to develop markers for DNA-based sex identification in wolverines using non-invasive samples. We developed two Y-linked markers, one that was specific to wolverine and one that also successfully identified sex in another mustelid. Importantly, none of the markers amplified potential prey species such as reindeer or rodents.     

Effect of niacin supplementation on digestibility, nitrogen utilisation and milk and blood variables in lactating dairy cows fed a diet with a negative rumen nitrogen balance

The aim of the present experiment was to determine if a niacin supplementation of 6 g/d to lactating dairy cow diets can compensate negative effects of a rumen nitrogen balance (RNB) deficit. A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows were successively assigned to one of three diets consisting of 10 kg maize silage (dry matter [DM] basis) and 7 kg DM concentrate: Diet RNB- (n = 6) with energy and utilisable crude protein at the duodenum (uCP) according to the average requirement of the animals but with a negative RNB (-0.41 g N/MJ metabolisable energy [ME]); Diet RNB0 (n = 7) with energy, uCP and a RNB (0.08 g N/MJ ME) according to the average requirement of the animals and, finally, Diet NA (n = 5), which was the same diet as RNB-, but supplemented with 6 g niacin/d. Samples of milk were taken on two consecutive days, blood samples were taken on one day pre- and post-feeding and faeces and urine were collected completely over five consecutive days. The negative RNB reduced milk and blood urea content and apparent total tract digestibility of DM, organic matter (OM) and neutral detergent fibre (NDF). Also N excretion with urine, the total N excreted with urine and faeces and the N balance were reduced when the RNB was negative. Supplementation of niacin elevated plasma glucose concentration after feeding and the N balance increased. Supplementing the diet with a negative RNB with niacin led to a more efficient use of dietary N thereby avoiding the negative effects of the negative RNB on the digestibility of DM, OM and NDF.     

Effect of roughage type and concentrate supplementation on follicle numbers and in vitro fertilisation and development of oocytes recovered from beef heifers

Increasing dietary energy tends to decrease the ovulatory response and produce fewer viable embryos following superovulation of beef cattle. Data in sheep indicate that high energy intake can decrease progesterone concentrations (P4), although effects in cattle are not as clear. The objectives were to evaluate the effects of roughage type and concentrate supplementation on P4 concentrations, follicle growth and subsequent oocyte fertilisation and embryo development in vitro. Forty-two beef heifers were allocated to 3 treatment groups: (i) silage ad libitum plus 6 kg concentrates (silage + conc.; n = 14); (ii) silage ad libitum (silage; n = 14) or (iii) hay ad libitum (hay; n = 14) for 40 days. Oestrus was synchronised using a controlled intravaginal progesterone releasing device (CIDR) for 7 days plus prostaglandin F2 alpha (15 mg luprostiol) administered 2 days before CIDR withdrawal. Ovaries were stimulated with 600 i.u. of follicle stimulating hormone (pFSH) administered in 6 equal doses at 12-h intervals, starting 12 days after CIDR withdrawal. Daily blood samples were collected from 3 days after CIDR insertion until CIDR withdrawal, and for another 3 days prior to pFSH, for P4 determination. Oocytes were recovered postmortem 12 h after the last pFSH injection, matured, fertilised and cultured in vitro. There was no overall effect of diet (P > 0.05) on P4 concentrations. The number of follicles grown in heifers on silage + conc (18.8 +/- 3.3), silage (23.5 +/- 3.4) or hay (18.1 +/- 2.6) were not affected by the dietary treatment (P > 0.05). The percentage of oocytes fertilised from heifers on hay (88%) was higher compared to oocytes from heifers on silage (79%; P < 0.05), but was not different (P > 0.05) compared to the proportion of oocytes from heifers on silage + conc. (86%). The percentage of fertilised oocytes that cleaved was higher from heifers on silage (94%; P < 0.01) compared with oocytes from heifers on hay (82%) or silage + conc. (86%). The proportion of embryos that developed to blastocyst was not different (P > 0.05) between groups of oocytes from heifers on silage + conc. (8%), silage (14%) or hay (15%). Heifers on silage produced numerically more blastocysts (silage: 19 from 14 heifers; silage + conc.: 8 from 14 heifers; hay: 12 from 14 heifers). These results suggest that dietary treatment used prior to oocyte recovery did not significantly influence the developmental competence of the oocytes in vitro.     

Development of a metabolomic approach based on urine samples and direct infusion mass spectrometry

The analysis of urine by direct infusion mass spectrometry suffers from ion suppression due to its high salt content and inter-sample variability caused by the differences in urine volume between persons. Thus, urine metabolomics requires a careful selection of the sample preparation procedure and a normalization strategy to deal with these problems. Several approaches were tested for metabolomic analysis of urine samples by direct infusion electrospray mass spectrometry (DI–ESI–MS), including solid phase extraction, liquid–liquid extraction, and sample dilution. In addition, normalization of results based on conductivity values and statistical treatment was performed to minimize sample variability. Both urine dilution and solid phase extraction with mixed mode sorbent considerably reduced the salt content in urine, providing comprehensive metabolomic fingerprints. Moreover, statistical data normalization enabled the correction of inter-sample physiological variability, improving the quality of results obtained. Therefore, high-throughput DI–ESI–MS fingerprinting of urine samples can be achieved with simple pretreatment procedures allowing the use of this noninvasive sampling in metabolomics. Finally, the optimized approach was tested in a pilot metabolomic investigation of urine samples from transgenic mice models of Alzheimer’s disease (APP/PS1) in order to illustrate the potential of the methodology.     

Impacts of protein supplementation during late gestation of beef cows on maternal skeletal muscle and liver tissues metabolism

Since nutritional requirements are increased at the end of gestation to meet the demands of the pregnant uterus, pregnant beef cows are susceptible to mobilization of body reserves (mainly fat and amino acids (AAs)) and to alter the metabolism of nutrients in the liver and muscle to support such demands. The objective of this study was to evaluate the effect of CP supplementation on maternal nutrient metabolism in the late gestation of beef cows grazing a low-quality pasture. Forty-three pregnant Nellore cows gestating male fetuses (average age = 6 years; average weight = 544 kg) at 193 ± 30 (mean ± SD) days (d) of gestation were divided into eight groups (experimental units, with four to five cows each). Treatments were (1) control (CON, n = 4): pasture-based (PB) diet without CP supplementation and (2) supplemented (SUP, n = 4): PB diet daily supplemented with 2 g/kg of BW of a 43.5% CP supplement. Liver and skeletal muscle biopsies were performed at 265 days of gestation and samples were collected for mRNA expression. On day 280 of gestation, blood samples were collected to assess plasma levels of AA. The CON-fed cows tended to have greater (P = 0.057) total circulating AA than SUP-fed cows. The circulating glycogenic AA was greater (P = 0.035) in CON than in SUP cows. CON cows was greater for histidine (P = 0.015), methionine (P = 0.007) and alanine (P = 0.036) than SUP cows. The CON- and SUP-fed showed no differences for gluconeogenesis, fatty acid transport and signaling axis markers in the liver. The mRNA expression of markers for skeletal muscle synthesis, p7056k (P = 0.060) and GSK3B (P = 0.096), tended to be greater in cows from CON than SUP group. No differences were found for mRNA expression of markers for skeletal muscle degradation. We conclude that CP supplementation to CP-restricted late-pregnant beef cows reduces the maternal tissue mobilization and changes the profile of plasma circulating AA and the mRNA expression of markers for the synthesis of skeletal muscle tissue.