Production and nitrogen metabolism in lactating dairy cows fed finely ground field pea plus soybean meal or canola meal with or without rumen-protected methionine supplementation.

We showed previously that dairy cows fed [diet dry matter (DM) basis] 25% finely ground field pea (GFP) plus rumen-protected (RP)-Met and RP-Lys had greater milk true protein yield and plasma Met concentration, but lower plasma His, compared with those fed GFP without rumen-protected AA supplementation. The goal of the present study was to investigate the effects of diets containing soybean meal (SBM) or canola meal (CM) with or without a source of RP-Met on production, nutrient digestibility, and N metabolism in cows fed 25% GFP. Sixteen mid-lactation Holstein cows were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments (21-d periods). Cows were fed (DM basis) 35% corn silage, 14% grass-legume haylage, 25% GFP, and 11% SBM or 13.5% CM with or without 0.095% RP-Met supplementation as Smartamine M (Adisseo USA Inc., Alpharetta, GA). Protein source effects were observed for most production variables; cows fed diets containing CM plus GFP had greater DM intake, yields of milk and milk fat and true protein, and milk N efficiency than those offered SBM plus GFP. Feeding CM plus GFP also reduced the concentrations of milk urea N and plasma urea N, and the urinary excretion of NH3 N and urea N, suggesting improved N use efficiency. Moreover, replacing SBM with CM increased the concentrations of all essential AA (except Arg) in plasma collected from the coccygeal blood vessels. A protein source × RP-Met interaction was observed for the concentration of His in coccygeal plasma, with circulating His decreasing only when RP-Met was supplemented to the diet containing SBM plus GFP. Based on the arteriovenous difference method, Lys was the first limiting AA overall, with Met being the first limiting AA in diets that did not receive RP-Met. Apparent total-tract digestibilities of DM, organic matter, N, and acid detergent fiber increased with feeding SBM plus GFP versus CM plus GFP. Most variables were not affected by RP-Met supplementation, except plasma Met concentration, which increased by 63%. Collectively, our results indicate that CM appears to be a better companion rumen-degradable protein source to GFP than SBM, due to improved yields of milk and milk protein and N use efficiency.

Production, milk iodine, and nutrient utilization in Jersey cows supplemented with the brown seaweed Ascophyllum nodosum (kelp meal) during the grazing season.

Kelp meal (KM) is a supplement made from the brown seaweed Ascophyllum nodosum, known to bioaccumulate iodine (I) and to be the richest source of phlorotannins, which can inhibit ruminal proteolysis and microbial growth. The objective of this study was to investigate the effects of KM on production, milk I, concentrations of blood metabolites, apparent total-tract digestibility of nutrients, and CH4 emissions in grazing dairy cows. Eight multiparous Jersey cows averaging (mean ± SD) 175 ± 60 d in milk and 12 primiparous Jersey cows averaging 142 ± 47 d in milk at the beginning of the study were assigned to either 0 g/d of KM (control diet, CTRL) or 113 g/d of KM (brown seaweed diet, BSW) in a randomized complete block design. Diets were formulated to yield a 70:30 forage-to-concentrate ratio and consisted of (dry matter basis): 48% cool-season perennial herbage and 52% partial TMR (pTMR). Each experimental period (n = 3) lasted 28 d, with data and sample collection taking place during the last 7 d of each period. Cows had approximately 16.5 h of access to pasture daily. Herbage dry matter intake increased, and total dry matter intake tended to increase in cows fed BSW versus the CTRL diet. Milk yield and concentrations and yields of milk components were not affected by diets. Similarly, blood concentrations of cortisol, glucose, fatty acids, and thyroxine did not change with feeding CTRL or BSW. However, a diet × period interaction was observed for milk I concentration; cows offered the BSW diet had greater milk I concentration during periods 1, 2, and 3, but the largest difference between BSW and CTRL was observed in period 2 (579 vs. 111 µg/L, respectively). Except for period 2, the concentration of milk I in cows fed KM did not exceed the 500 µg/L threshold recommended for human consumption. Diet × period interactions were also found for serum triiodothyronine concentration, total-tract digestibilities of crude protein and acid detergent fiber, CH4 production, and urinary excretion of purine derivatives. Overall, the lack of KM effects on milk yield and concentrations and yields of milk components indicate that dairy producers should consider costs before making KM supplementation decisions during the grazing season. Future research is needed to evaluate the concentration of I in retail organic milk because of the high prevalence of KM supplementation in northeastern and midwestern US organic dairies and possibly in other regions of the country.

Production, milk fatty acid profile, and nutrient utilization in grazing dairy cows supplemented with ground flaxseed.

Flaxseed has been extensively used as a supplement for dairy cows because of its high concentrations of energy and the n-3 fatty acid (FA) cis-9,cis-12,cis-15 18:3. However, limited information is available regarding the effect of ground flaxseed on dry matter intake (DMI), ruminal fermentation, and nutrient utilization in grazing dairy cows. Twenty multiparous Jersey cows averaging (mean ± standard deviation) 111 ± 49 d in milk in the beginning of the study were used in a randomized complete block design to investigate the effects of supplementing herbage (i.e., grazed forage) with ground corn-soybean meal mix (control diet = CTRL) or ground flaxseed (flaxseed diet = FLX) on animal production, milk FA, ruminal metabolism, and nutrient digestibility. The study was conducted from June to September 2013, with data and sample collection taking place on wk 4, 8, 12, and 16. Cows were fed a diet formulated to yield a 60:40 forage-to-concentrate ratio consisting of (dry matter basis): 40% cool-season perennial herbage, 50% partial total mixed ration, and 10% of ground corn-soybean meal mix or 10% ground flaxseed. However, estimated herbage DMI averaged 5.59 kg/d or 34% of the total DMI. Significant treatment by week interactions were observed for milk and blood urea N, and several milk FA (e.g., trans-10 18:1). No significant differences between treatments were observed for herbage and total DMI, milk yield, feed efficiency, concentrations and yields of milk components, and urinary excretion of purine derivatives. Total-tract digestibility of organic matter decreased, whereas that of neutral detergent fiber increased with feeding FLX versus CTRL. No treatment effects were observed for ruminal concentrations of total volatile FA and NH3-N, and ruminal proportions of acetate and propionate. Ruminal butyrate tended to decrease, and the acetate-to-propionate ratio decreased in the FLX diet. Most saturated and unsaturated FA in milk fat were changed. Specifically, milk proportion of cis-9,cis-12,cis-15 18:3, Σn-3 FA, and Σ18C FA increased, whereas that of cis-9,cis-12 18:2, Σn-6 FA, Σ odd-chain FA, Σ<16C FA, and Σ16C FA decreased with feeding FLX versus the CTRL diet. In conclusion, feeding FLX did not change yields of milk and milk components, but increased milk n-3 FA. Therefore, costs and industry adoption of premiums for n-3-enriched milk will determine the adoption of ground flaxseed in pasture-based dairy farms.

Production and nitrogen utilization in lactating dairy cows fed ground field peas with or without ruminally protected lysine and methionine.

Previous research has shown that cows fed ≥24% of the diet dry matter (DM) as field peas decreased milk yield as well as concentration and yield of milk protein, possibly due to reduced DM intake and limited supply of Lys and Met. Twelve multiparous and 4 primiparous lactating Holstein cows were randomly assigned to 1 of 4 diets in a replicated 4 × 4 Latin square design. The diets contained (DM basis) 34.8% corn silage, 15.2% grass-legume silage, 5.9% roasted soybean, 2.4% mineral-vitamin premix, 2.0% alfalfa pellets, and either (1) 36% ground corn, 2.4% soybean meal, and 1.3% urea (UR), (2) 29.7% ground corn, 9.8% soybean meal, 0.13% ruminally protected (RP) Lys, and 0.07% RP-Met (CSBAA), (3) 25% ground field peas, 12.3% ground corn, and 2.4% soybean meal (FP), or (4) FP supplemented with 0.15% RP-Lys and 0.05% RP-Met (FPAA). Our objective was to test the effects of FP versus UR, FPAA versus CSBAA, and FPAA versus FP on milk yield and composition, N utilization, nutrient digestibility, ruminal fermentation profile, and plasma concentration of AA. Milk yield did not differ across diets. Compared with cows fed UR, those fed FP had greater DM intake, concentration and yield of milk true protein, apparent total-tract digestibility of fiber, urinary excretion of purine derivatives, and concentrations of total volatile fatty acids in the rumen and Lys in plasma, and less milk urea N and ruminal NH3-N. The concentration of milk urea N, as well as the concentration and yield of milk fat increased in cows fed FPAA versus CSBAA. Moreover, cows fed FPAA had greater ruminal concentration of total volatile fatty acids, increased proportions of acetate and isobutyrate, and decreased proportions of propionate and valerate than those fed CSBAA. The plasma concentrations of His, Leu, and Phe decreased, whereas plasma Met increased and plasma Lys tended to increase in cows fed FPAA versus CSBAA. Concentration of milk true protein, but not yield, was increased in cows fed FPAA versus FP. However, cows fed FPAA showed decreased concentrations of His and Leu in plasma compared with those fed FP. Overall, compared with the CSBAA diet, feeding FPAA did not negatively affect milk yield and milk protein synthesis. Furthermore, RP-Lys and RP-Met supplementation of the FP diet did not improve milk yield or milk protein synthesis, but decreased urinary urea N excretion.

Geospatial variability of soil CO2-C exchange in the main terrestrial ecosystems of Keller Peninsula, Maritime Antarctica.

Soils and vegetation play an important role in the carbon exchange in Maritime Antarctica but little is known on the spatial variability of carbon processes in Antarctic terrestrial environments. The objective of the current study was to investigate (i) the soil development and (ii) spatial variability of ecosystem respiration (ER), net ecosystem CO2 exchange (NEE), gross primary production (GPP), soil temperature (ST) and soil moisture (SM) under four distinct vegetation types and a bare soil in Keller Peninsula, King George Island, Maritime Antarctica, as follows: site 1: moss-turf community; site 2: moss-carpet community; site 3: phanerogamic antarctic community; site 4: moss-carpet community (predominantly colonized by Sanionia uncinata); site 5: bare soil. Soils were sampled at different layers. A regular 40-point (5×8 m) grid, with a minimum separation distance of 1m, was installed at each site to quantify the spatial variability of carbon exchange, soil moisture and temperature. Vegetation characteristics showed closer relation with soil development across the studied sites. ER reached 2.26µmolCO2m(-2)s(-1) in site 3, where ST was higher (7.53°C). A greater sink effect was revealed in site 4 (net uptake of 1.54µmolCO2m(-2)s(-1)) associated with higher SM (0.32m(3)m(-3)). Spherical models were fitted to describe all experimental semivariograms. Results indicate that ST and SM are directly related to the spatial variability of CO2 exchange. Heterogeneous vegetation patches showed smaller range values. Overall, poorly drained terrestrial ecosystems act as CO2 sink. Conversely, where ER is more pronounced, they are associated with intense soil carbon mineralization. The formations of new ice-free areas, depending on the local soil drainage condition, have an important effect on CO2 exchange. With increasing ice/snow melting, and resulting widespread waterlogging, increasing CO2 sink in terrestrial ecosystems is expected for Maritime Antarctica.

Prepartum supplementation of nicotinic acid: Effects on health of the dam, colostrum quality, and acquisition of immunity in the calf.

Nicotinic acid (NA) has been shown to reduce lipolysis, alter milk components and the ruminal environment, and increase blood flow. Increased blood flow to the mammary gland during colostrogenesis might increase nutrients and immunoglobulin concentration of colostrum. Twenty-six multiparous Holstein cows were housed in a tiestall barn. Cows were blocked by expected calving date and randomly assigned to 1 of 2 treatments 4 wk prepartum: (1) 0g/d of NA (control, CON) or (2) 48g/d of NA (NA). Total mixed ration amounts fed and refused were measured daily to determine dry matter intake. Blood samples were collected from dams every Monday, Wednesday, and Friday from the coccygeal vein or artery and were analyzed for glucose, nonesterified fatty acids (NEFA), and ß-hydroxybutyrate (BHB). Colostrum was collected and weighed within 90 min of parturition. Colostral immunoglobulin G (IgG) concentration was analyzed using radial immunodiffusion assay. Calves were removed from their dams before suckling and weighed within 30 min after birth. Calves received 3 L of a lacteal-based colostrum replacer that provided a total of 225.8g of IgG within 2h of birth. Calf blood samples were collected via jugular venipuncture at 0 and 24h of age and analyzed for IgG concentration and determination of apparent efficiency of absorption. Colostrum yield, dry matter intake, IgG yield, and fat and solids percentage of colostrum did not differ between treatments. Serum concentrations of glucose and BHB were not affected by treatment. We detected an effect of week on serum glucose concentrations at calving and on serum BHB concentrations at 1 wk postpartum. There was a treatment by week effect for serum NEFA concentrations at 1 wk postpartum, where cows that received NA prepartum had higher serum NEFA concentration than CON cows, indicating that a NEFA rebound occurred. No differences were observed for calf body weight, 0- or 24-h serum IgG concentration, or apparent efficiency of absorption. Supplementation of NA increased IgG concentration in colostrum from 73.8 to 86.8g/L. Results indicate that 48g/d of supplemental NA during the prepartum period improved colostrum quality.

Integrating spot short-term measurements of carbon emissions and backward dietary energy partition calculations to estimate intake in lactating dairy cows fed ad libitum or restricted.

The objective of this study was to use spot short-term measurements of CH4 (QCH4) and CO2 (QCO2) integrated with backward dietary energy partition calculations to estimate dry matter intake (DMI) in lactating dairy cows. Twelve multiparous cows averaging 173±37d in milk and 4 primiparous cows averaging 179±27d in milk were blocked by days in milk, parity, and DMI (as a percentage of body weight) and, within each block, randomly assigned to 1 of 2 treatments: ad libitum intake (AL) or restricted intake (RI=90% DMI) according to a crossover design. Each experimental period lasted 22d with 14d for treatments adaptation and 8d for data and sample collection. Diets contained (dry matter basis): 40% corn silage, 12% grass-legume haylage, and 48% concentrate. Spot short-term gas measurements were taken in 5-min sampling periods from 15 cows (1 cow refused sampling) using a portable, automated, open-circuit gas quantification system (GreenFeed, C-Lock Inc., Rapid City, SD) with intervals of 12h between the 2daily samples. Sampling points were advanced 2h from a day to the next to yield 16 gas samples per cow over 8d to account for diurnal variation in QCH4 and QCO2. The following equations were used sequentially to estimate DMI: (1) heat production (MJ/d)=(4.96 + 16.07 ÷ respiratory quotient) × QCO2; respiratory quotient=0.95; (2) metabolizable energy intake (MJ/d)=(heat production + milk energy) ± tissue energy balance; (3) digestible energy (DE) intake (MJ/d)=metabolizable energy + CH4 energy + urinary energy; (4) gross energy (GE) intake (MJ/d)=DE + [(DE ÷ in vitro true dry matter digestibility) - DE]; and (5) DMI (kg/d)=GE intake estimated ÷ diet GE concentration. Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC) and Fit Model procedure in JMP (α=0.05; SAS Institute Inc.). Cows significantly differed in DMI measured (23.8 vs. 22.4kg/d for AL and RI, respectively). Dry matter intake estimated using QCH4 and QCO2 coupled with dietary backward energy partition calculations (Equations 1 to 5 above) was highest in cows fed for AL (22.5 vs. 20.2kg/d). The resulting R(2) were 0.28 between DMI measured and DMI estimated by gaseous measurements, and 0.36 between DMI measured and DMI predicted by the National Research Council model (2001). Results showed that spot short-term measurements of QCH4 and QCO2 coupled with dietary backward estimations of energy partition underestimated DMI by 7.8%. However, the approach proposed herein was able to significantly discriminate differences in DMI between cows fed for AL or RI.

Short communication: Substituting dry distillers grains with solubles and rumen-protected amino acids for soybean meal in late-lactation cows' diets based on corn silage or ryegrass silage.

Excess protein in dairy cattle diets increases production costs and contributes to environmental pollution. The objective of the present study was to evaluate the effect of feeding dry distillers grains with solubles (DDGS) supplemented with rumen-protected Lys and Met in place of solvent-extracted soybean meal on the performance of late-lactation cows. Two experiments were carried out, with each using 24 late-lactating dairy cows distributed among 4 pens. In trial 1, corn silage was the main forage source. Control (HP1) total mixed ration (TMR) contained 16.3% crude protein (CP) with soybean meal as the main protein source. Treatment TMR (LP1) had 13.7% CP when soybean meal was replaced with DDGS and rumen-protected Lys and Met. Forage in trial 2 was ryegrass silage; control TMR (HP2; 15.4% CP) contained soybean meal and rumen-protected Met, whereas treatment TMR (LP2; 13.8% CP) contained DDGS and rumen-protected Lys and Met. Trials were analyzed as crossover design using the MIXED procedure of SAS (SAS Institute Inc., Cary NC) with cow as sampling unit and pen as the experimental unit. Treatments were similar in dry matter intake (21.0 and 20.4 kg/cow per day for HP1 and LP1, respectively) and milk yield (20.7 and 20.5 kg/cow per day for HP1 and LP1, respectively) during trial 1. Milk composition was similar between treatments, averaging 4.22, 3.73, 4.54, and 9.15, respectively, for fat, protein, lactose, and solids nonfat. Milk urea nitrogen decreased from 17.2 mg/dL for HP1 to 9.93 mg/dL for LP1. In trial 2, no significant differences were observed for dry matter intake (21.4 and 20.9 kg/cow per day for HP2 and LP2, respectively), milk yield (28.1 and 26.6 kg/d for HP2 and LP2, respectively), fat yield (0.99 vs. 0.92 kg/d for HP2 and LP2, respectively), protein yield (0.94 vs. 0.86 kg/d for HP2 and LP2, respectively) and lactose yield (1.37 vs. 1.28 for HP2 and LP2, respectively). Milk urea nitrogen decreased from 9.88 mg/dL with HP2 to 6.39 mg/dL with the LP2 treatment. Milk N efficiency tended to be higher for LP treatments in trial 1, but not in trial 2. Low milk urea N suggested nitrogen losses to the environment may be lower when cows were fed diets based on DDGS in both trials. The studies indicated that DDGS with rumen-protected Lys and Met could substitute solvent-extracted soybean meal in low-protein corn silage- and ryegrass silage-based diets for late-lactation dairy cows averaging 20.6 or 27.4 kg of milk/d, respectively.

Incremental amounts of ground flaxseed decrease milk yield but increase n-3 fatty acids and conjugated linoleic acids in dairy cows fed high-forage diets(1).

The objective of this study was to investigate the effect of incremental amounts of ground flaxseed (GFX) on milk yield and concentrations and yields of milk components, milk fatty acids (FA) profile, ruminal metabolism, and nutrient digestibility in dairy cows fed high-forage diets. Twelve multiparous Jersey cows averaging (mean ± SD) 112±68d in milk and 441±21kg of body weight and 8 primiparous Jersey cows averaging 98±43d in milk and 401±43kg of body weight were randomly assigned to treatment sequences in a replicated 4×4 Latin square design. Each period lasted 21d with 14d for diet adaptation and 7d for data and sample collection. Treatments were fed as a total mixed ration (63:37 forage-to-concentrate ratio) with corn meal and soybean meal replaced by incremental levels (i.e., 0, 5, 10, or 15% diet dry matter) of GFX. The ruminal molar proportions of acetate and butyrate decreased linearly with GFX supplementation, whereas the ruminal molar proportion of propionate increased linearly resulting in decreased acetate-to-propionate ratio. Apparent total-tract digestibilities of nutrients either decreased (dry matter) or tended to decrease (organic matter, neutral detergent fiber, acid detergent fiber) linearly in cows fed GFX. Milk yield decreased linearly in cows fed increasing amounts of GFX, which is explained by the linear reduction in dry matter intake. Except for the concentrations of milk protein and urea N, which decreased linearly with GFX supplementation, no other changes in the concentration of milk components were observed. However, yields of milk protein and fat decreased linearly with GFX supplementation. The linear decrease in the yields of milk fat and protein are explained by reduced milk yield, whereas that in milk urea N is explained by decreased crude protein intake. No treatment effects were observed for plasma urea N and nonesterified fatty acids, serum cortisol, and body weight change. Milk odd- and branched-chain FA and saturated FA decreased linearly with GFX supplementation. Milk trans-11 18:1, α-linolenic acid, cis-9,trans-11 18:2, and the sum of n-3 FA all increased linearly and quadratically, whereas the milk ratio of n-6 to n-3 decreased linearly in cows fed GFX. Overall, compared with the control diet (0% GFX), the diet with 15% GFX supplementation resulted in the lowest milk yield but highest milk proportions and yields (data not shown) of cis-9,trans-11 18:2 and n-3 FA.

Short communication: Use of a portable, automated, open-circuit gas quantification system and the sulfur hexafluoride tracer technique for measuring enteric methane emissions in Holstein cows fed ad libitum or restricted.

The objective of this study was to measure enteric CH4 emissions using a new portable automated open-circuit gas quantification system (GQS) and the sulfur hexafluoride tracer technique (SF6) in midlactation Holstein cows housed in a tiestall barn. Sixteen cows averaging 176 ± 34 d in milk, 40.7 ± 6.1 kg of milk yield, and 685 ± 49 kg of body weight were randomly assigned to 1 out of 2 treatments according to a crossover design. Treatments were (1) ad libitum (adjusted daily to yield 10% orts) and (2) restricted feed intake [set to restrict feed by 10% of baseline dry matter intake (DMI)]. Each experimental period lasted 22d, with 14 d for treatment adaptation and 8d for data and sample collection. A common diet was fed to the cows as a total mixed ration and contained 40.4% corn silage, 11.2% grass-legume haylage, and 48.4% concentrate on a dry matter basis. Spot 5-min measurements using the GQS were taken twice daily with a 12-h interval between sampling and sampling times advanced 2h daily to account for diurnal variation in CH4 emissions. Canisters for the SF6 method were sampled twice daily before milking with 4 local background gas canisters inside the barn analyzed for background gas concentrations. Enteric CH4 emissions were not affected by treatments and averaged 472 and 458 g/d (standard error of the mean = 18 g/d) for ad libitum and restricted intake treatments, respectively (data not shown). The GQS appears to be a reliable method because of the relatively low coefficients of variation (ranging from 14.1 to 22.4%) for CH4 emissions and a moderate relationship (coefficient of determination = 0.42) between CH4 emissions and DMI. The SF6 resulted in large coefficients of variation (ranging from 16.0 to 111%) for CH4 emissions and a poor relationship (coefficient of determination = 0.17) between CH4 emissions and DMI, likely because of limited barn ventilation and high background gas concentration. Research with improved barn ventilation systems or outdoors is warranted to further assess the GQS and SF6 methodologies.

Interactions of corn meal or molasses with a soybean-sunflower meal mix or flaxseed meal on production, milk fatty acid composition, and nutrient utilization in dairy cows fed grass hay-based diets.

We investigated the interactions of corn meal or molasses [nonstructural carbohydrate (NSC) supplements] with a soybean-sunflower meal mix or flaxseed meal [rumen-degradable protein (RDP) supplements] on animal production, milk fatty acids profile, and nutrient utilization in dairy cows fed grass hay diets. Eight multiparous and 8 primiparous Jersey cows averaging 135±49d in milk and 386±61kg of body weight in the beginning of the study were randomly assigned to 4 replicated 4×4 Latin squares with a 2×2 factorial arrangement of treatments. Each period lasted 19d with 14d for diet adaptation and 5d for data and sample collection. Cows were fed diets composed of mixed-mostly grass hay plus 1 of the following 4 concentrate blends: (1) corn meal plus a protein mix containing soybean meal and sunflower meal; (2) corn meal plus flaxseed meal; (3) liquid molasses plus a protein mix containing soybean meal and sunflower meal; or (4) liquid molasses plus flaxseed meal. Data were analyzed for main effects of NSC and RDP supplements, and the NSC × RDP supplement interactions. Significant NSC × RDP supplement interactions were observed for milk urea N, milk N efficiency, and the sums of milk saturated, monounsaturated, and polyunsaturated fatty acids. No effect of NSC supplements was observed for nutrient intake and milk yield. However, 4% fat-corrected milk (-0.70kg/d) and energy-corrected milk (-0.60kg/d) were significantly reduced in cows fed liquid molasses due to a trend to decreased concentration of milk fat (-0.17%). Diets with liquid molasses resulted in increased (+35%) concentration and yield of milk enterolactone, indicating that this mammalian lignan can be modulated by supplements with different NSC profiles. Overall, NSC and RDP supplements profoundly changed the milk fatty acid profile, likely because of differences in fatty acids intake, Δ(9)-desaturase indices, and ruminal biohydrogenation pathways. Feeding liquid molasses significantly reduced plasma urea N (-1.2mg/dL), urinary N excretion (-20g/d), and N digestibility (-3.2 percentage units). Flaxseed meal significantly reduced yields of milk (-1.3kg/d), milk fat (-90g/d), and milk lactose (-60g/d), but significantly increased the concentration and yield of milk enterolactone. Further research is needed to elucidate the negative responses of flaxseed meal on yields of milk and milk components.

Kidney transplantation: evaluation and clinical outcome of 237 recipients at low, medium, high, or strong immunological risk of rejection.

BACKGROUND: Donor-specific antibodies (DSAs) play a fundamental role in kidney transplantation. The identification of DSAs is an essential rejection parameter. PATIENTS AND METHODS: We evaluated a protocol in 237 patients receiving kidneys from living (LDs) and deceased donors (DDs). Recipients were classified as being at low (LR), medium (MR), high (HR), or strong (SR) risk of rejection based on Luminex panel reactive antibody (PRA)-single antigen beads (SABs). Grafts that survived for 1 year were evaluated. RESULTS: Of the 237 transplanted patients, 129 (54.43%) received a kidney from an LD and 108 (45.57%) from a DD. Of 95 LR recipients receiving kidneys from LDs, 2 patients lost the graft due to non-immunological causes. Of 34 MR recipients, 13 had rejection episodes, and 2 lost the graft by AMR and one by cellular rejection (CR). Of 108 recipients receiving a kidney from a DD, 59 (54.63%) were LR, 31 (28.70%) MR, 11 (10.19%) HR, and 7 (6.48%) SR. Twenty of all transplanted recipients lost their grafts; 4 were due to clinical causes, 4 by cellular rejection, and 12 by antibody-mediated rejection (AMR) with PRA-SAB mean fluorescent intensity of 530 to 12,591. One-year graft survival for LD transplanted LR and MR patients was 97.6% and 94.1%, respectively (P = .004). In DD recipients, the LR vs MR SD was P = .011, and for LR vs HR + SR it was P = .001. For MR vs HR+SR no SD was found (P = .323). CONCLUSION: Rejections were detected in 51 patients (21.52%). Graft failure occurred in 16 patients (6.75%). A total of 218 (91.98%) recipients maintained good kidney function after 1 year. This protocol based on fluxogram risk assessment of AMR provided fast and precise immunological evaluation of recipients and donors and stratification by immunological risk of AMR.

A Study on the Quality and Identity of Brazilian Pampa Biome Honey: Evidences for Its Beneficial Effects against Oxidative Stress and Hyperglycemia.

We characterized, for the first time, the quality and identity of Brazilian Pampa biome honey and its antioxidant properties in vitro (FRAP, DDPH and ABTS). The potential protective effect of honey against oxidative stress induced by iron (Fe) and paraquat, (PQ) in a Drosophila melanogaster model (in vivo) was also tested. The results indicated that all honey samples tested showed antioxidant activity in vitro. Flies treated with honey showed increased lifespan and were protected against oxidative stress induced by Fe and PQ. Despite the high concentration of sugars in honey (approximately 70-80%), our results demonstrate a hypoglycemic-like effect of honey in Drosophila. Thus, this study demonstrates the high quality of Brazilian Pampa biome honey as well as its significant antioxidant activity in vitro and in vivo, pointing to the potential use of this natural product as an alternative in the therapy of oxidative stress-associated diseases.

Effects of Hg(II) exposure on MAPK phosphorylation and antioxidant system in D. melanogaster.

The heavy metal mercury is a known toxin, but while the mechanisms involved in mercury toxicity have been well demonstrated in vertebrates, little is known about toxicological effects of this metal in invertebrates. Here, we present the results of our study investigating the effects associated with exposure of fruit fly Drosophila melanogaster to inorganic mercury (HgCl2 ). We quantify survival and locomotor performance as well as a variety of biochemical parameters including antioxidant status, MAPK phosphorylation and gene expression following mercury treatment. Our results demonstrate that exposure to Hg(II) through diet induced mortality and affected locomotor performance as evaluated by negative geotaxis, in D. melanogaster. We also saw a significant impact on the antioxidant system including an inhibition of acetylcholinesterase (Ache), glutathione S-transferase (GST) and superoxide dismutase (SOD) activities. We found no significant alteration in the levels of mRNA of antioxidant enzymes or NRF-2 transcriptional factor, but did detect a significant up regulation of the HSP83 gene. Mercury exposure also induced the phosphorylation of JNK and ERK, without altering p38(MAPK) and the concentration of these kinases. In parallel, Hg(II) induced PARP cleavage in a 89 kDa fragment, suggesting the triggering of apoptotic cell death in response to the treatment. Taken together, this data clarifies and extends our understanding of the molecular mechanisms mediating Hg(II) toxicity in an invertebrate model.

Microarray diagnosis of antibody-mediated rejection in kidney transplant biopsies: an international prospective study (INTERCOM).

In a reference set of 403 kidney transplant biopsies, we recently developed a microarray-based test that diagnoses antibody-mediated rejection (ABMR) by assigning an ABMR score. To validate the ABMR score and assess its potential impact on practice, we performed the present prospective INTERCOM study (clinicaltrials.gov NCT01299168) in 300 new biopsies (264 patients) from six centers: Baltimore, Barcelona, Edmonton, Hannover, Manchester and Minneapolis. We assigned ABMR scores using the classifier created in the reference set and compared it to conventional assessment as documented in the pathology reports. INTERCOM documented uncertainty in conventional assessment: In 41% of biopsies where ABMR features were noted, the recorded diagnoses did not mention ABMR. The ABMR score correlated with ABMR histologic lesions and donor-specific antibodies, but not with T cell-mediated rejection lesions. The agreement between ABMR scores and conventional assessment was identical to that in the reference set (accuracy 85%). The ABMR score was more strongly associated with failure than conventional assessment, and when the ABMR score and conventional assessment disagreed, only the ABMR score was associated with early progression to failure. INTERCOM confirms the need to reduce uncertainty in the diagnosis of ABMR, and demonstrates the potential of the ABMR score to impact practice.

Assessing the research and education needs of the organic dairy industry in the northeastern United States.

Demographic and management data about organic dairies have been reported previously, but the current study is the first needs assessment of research and educational priorities of organic dairy farmers in the northeastern United States based directly upon their input. Our objectives were to (1) develop an initial understanding of the emerging research and educational needs of organic dairy farmers in the northeastern United States via focus group interviews, and (2) prioritize the needs identified by the focus groups with a broader population of organic dairy farmers via survey methods. Focus group interviews determined the questions used for the survey questionnaire distributed to 1,200 members of the Northeast Organic Dairy Producers Alliance. The members were asked about demographic information, but more importantly, challenges concerning business management and marketing, organic certification, and animal nutrition, health, and reproduction. The results (183 respondents, 15% response rate) were parsed by region (New England farms compared with New York and Pennsylvania farms), herd size (i.e., 12 to 37, 38 to 59, and >60 cows), and years of organic certification (<4 yr vs. ≥ 4 yr); however, no differences between regions were observed for demographic data. The average farm consisted of 309 acres and 57 milking cows, on which most of the forage was homegrown but grains were purchased (73% of farms). Among the greatest challenges identified by the farmers were obtaining a steady, fair price for milk (85% respondents); determining dry matter intake for animals on pasture (76%); and controlling nuisance flies (89%). Needs for additional research included organic treatments for mastitis (92% respondents), growing forages for organic production (84%), and developing value-added products (84%). Farms with <4 yr of organic certification were concerned with level of knowledge and experience of local certifiers, whereas organic producers with ≥ 4 yr of organic certification were more interested in field testing of new organic products. Opportunities for educational programs included learning about direct marketing possibilities (76% respondents) and providing training to regional veterinarians interested in organic remedies (91%). In conclusion, the information obtained from the current needs assessment provides a foundation for future research proposals and educational outreach programs, germane to stakeholder needs, which could benefit the organic dairy industry within the region and beyond.

Potential impact of microarray diagnosis of T cell-mediated rejection in kidney transplants: The INTERCOM study.

We previously developed a microarray-based test for T cell-mediated rejection (TCMR) in a reference set of 403 biopsies. To determine the potential impact of this test in clinical practice, we undertook INTERCOM, a prospective international study of 300 indication biopsies from 264 patients (ClinicalTrials.gov NCT01299168). Biopsies from six centers-Baltimore, Barcelona, Edmonton, Hannover, Manchester and Minneapolis-were analyzed by microarrays, assigning TCMR scores by an algorithm developed in the reference set and comparing TCMR scores to local histology assessment. The TCMR score correlated with histologic TCMR lesions-tubulitis and interstitial infiltration. The accuracy for primary histologic diagnoses (0.87) was similar to the reference set (0.89). The TCMR scores reclassified 77/300 biopsies (26%): 16 histologic TCMR were molecularly non-TCMR; 15 histologic non-TCMR were molecularly TCMR, including 6 with polyoma virus nephropathy; and all 46 "borderline" biopsies were reclassified as TCMR (8) or non-TCMR (38). Like the reference set, discrepancies were primarily in situations where histology has known limitations, for example, in biopsies with scarring and inflammation/tubulitis potentially from other diseases. Neither the TCMR score nor histologic TCMR was associated with graft loss. Thus the molecular TCMR score has potential to add new insight, particularly in situations where histology is ambiguous or potentially misleading.

Is the lobster cockroach Nauphoeta cinerea a valuable model for evaluating mercury induced oxidative stress?

Organic and inorganic forms of mercury are highly neurotoxic environmental contaminants. The exact mechanisms involved in mercury neurotoxicity are still unclear. Oxidative stress appears to play central role in this process. In this study, we aimed to validate an insect-based model for the investigation of oxidative stress during mercury poisoning of lobster cockroach Nauphoeta cinerea. The advantages of using insects in basic toxicological studies include the easier handling, rapid proliferation/growing and absence of ethical issues, comparing to rodent-based models. Insects received solutions of HgCl2 (10, 20 and 40mgL(-1) in drinking water) for 7d. 24h after mercury exposure, animals were euthanized and head tissue samples were prepared for oxidative stress related biochemical determinations. Mercury exposure caused a concentration dependent decrease in survival rate. Cholinesterase activity was unchanged. Catalase activity was substantially impaired after mercury treatment 40mgL(-1). Likewise, GST had a significant decrease, comparing to control. Peroxidase and thioredoxin reductase activity was inhibited at concentrations of 20mgL(-1) and 40mgL(-1) comparing to control. These results were accompanied by decreased GSH levels and increased hydroperoxide and TBARS formation. In conclusion, our results show that mercuric compounds are able to induce oxidative stress signs in insect by modulating survival rate as well as inducing impairments on important antioxidant systems. In addition, our data demonstrates for the first time that Nauphoeta cinerea represents an interesting animal model to investigate mercury toxicity and indicates that the GSH and thioredoxin antioxidant systems plays central role in Hg induced toxicity in insects.

Urinary MCP-1 and RBP: independent predictors of renal outcome in macroalbuminuric diabetic nephropathy.

BACKGROUND: Albuminuria has been considered a sine qua non condition for the diagnosis of diabetic nephropathy (DN) and has been widely used as a surrogate outcome of chronic kidney disease (CKD). However, recent data suggest that albuminuria may fail as a biomarker in a subset of patients, and the search for novel markers is intense. METHODS: We analyzed the role of urinary RBP and of serum and urinary cytokines (TGF-beta, MCP-1 and VEGF) as predictors of the risk of dialysis, doubling of serum creatinine or death (primary outcome, PO) in 56 type 2 diabetic patients with macroalbuminuric DN. RESULTS: Mean follow-up time was 30.7±10 months. Urinary RBP and MCP-1 were significantly higher in patients presenting the PO, whereas no difference was shown for TGF-ß or VEGF. In the Cox regression, urinary RBP, MCP-1 and VEGF were positively associated and serum VEGF was inversely related to the risk of the PO. However, after adjustments for creatinine clearance, proteinuria, and blood pressure only urinary RBP (OR 11.6; 95% CI 2.7-49.2, p=0.001 for log RBP) and urinary MCP-1 (OR 11.0; 95% CI 1.6-76.4, p=0.02 for log MCP-1) remained as significant independent predictors of the PO. CONCLUSION: Urinary RBP and MCP-1 are independently related to the risk of CKD progression in patients with macroalbuminuric DN. Whether these biomarkers have a role in the setting of normoalbuminuria and microalbuminuria in DN should be further investigated.

Evidences for a role of glutathione peroxidase 4 (GPx4) in methylmercury induced neurotoxicity in vivo.

We evaluated the activity and expression of antioxidant enzymes in the cerebellum and cortex of Swiss adult male mice exposed to methylmercury (MeHg) in drinking water (40mg/L) during 21 days. The activity of glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were determined spectrophotometrically. The expression (protein levels) of GPx1 and GPx4 isoforms, TrxR1 as well as heat shock protein 70 (HSP70) were evaluated using specific antibodies and normalized by actin levels. The exposure of mice to MeHg caused a significant impairment in locomotors performance in the open field test (crossings and rearing). This result was followed by a significant reduction of GPx and TrxR activities in the cerebellum and cortex when compared to untreated animals. We also observed a substantial decrease in GPx1, GPx4 and TrxR1 protein levels in the cerebellum, while in the cerebral cortex, only GPx4 and TrxR1 were decreased after MeHg treatment. The activities of the antioxidant enzymes GR, GST, CAT and SOD were increased in the cerebellum after MeHg administration to mice. In contrast, only CAT was increased in the cerebral cortex of MeHg-treated animals. The expression of HSP70 was up-regulated only in the cerebellum where MeHg-exposed mice showed a significant increase in the immunocontent of HSP70 when compared to controls. This is the first report showing a role for GPx4 in the neurotoxicity induced by MeHg in vivo. In addition, our data indicates that the selenoproteins GPx and TrxR as main targets during MeHg exposure, which may be considered in biomarker studies.